LOCUS       pTRAST_GG               6871 bp    DNA     circular SYN 29-JUN-2026
DEFINITION  Single-vector trastuzumab IgG1 mAb expression vector for CHO-K1
            (BsaI Golden Gate).
ACCESSION   pTRAST_GG
VERSION     pTRAST_GG
KEYWORDS    .
SOURCE      .
  ORGANISM  synthetic construct
            .
FEATURES             Location/Qualifiers
     promoter        1..584
                     /label="CMV enhancer/promoter"
     intron          585..717
                     /label="chimeric intron"
     misc_feature    718..721
                     /label="fusion site J1 (AGGA)"
     misc_feature    722..727
                     /label="Kozak"
     CDS             728..2140
                     /label="heavy chain: Ig-kappa leader + trastuzumab VH +
                     human IgG1 (CHO-opt)"
                     /translation="METDTLLLWVLLLWVPGSTGEVQLVESGGGLVQPGGSLRLSCAAS
                     GFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMN
                     SLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA
                     ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
                     CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE
                     VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK
                     EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD
                     IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
                     YTQKSLSLSPGK"
                     /codon_start=1
     misc_feature    2141..2144
                     /label="fusion site J2 (TAGA)"
     misc_feature    2145..2733
                     /label="EMCV IRES"
     CDS             2731..3435
                     /label="light chain: Ig-kappa leader + trastuzumab VL +
                     kappa C (CHO-opt)"
                     /translation="METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASVGDRVTITCRA
                     SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFA
                     TYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE
                     AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
                     SPVTKSFNRGEC"
                     /codon_start=1
     misc_feature    3436..4024
                     /label="WPRE"
     polyA_signal    4025..4249
                     /label="bGH poly(A)"
     misc_feature    4250..4253
                     /label="fusion site J3 (GCGA)"
     promoter        4254..4583
                     /label="SV40 promoter"
     misc_feature    4584..4589
                     /label="Kozak"
     CDS             4590..5189
                     /label="puromycin N-acetyltransferase (pac, CHO-opt)"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
                     /codon_start=1
     polyA_signal    5190..5311
                     /label="SV40 poly(A)"
     misc_feature    5312..5315
                     /label="fusion site J4 (TGAA)"
     rep_origin      5316..5901
                     /label="pUC origin"
     promoter        5902..6006
                     /label="AmpR promoter"
     CDS             6007..6867
                     /label="AmpR (bla)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    6868..6871
                     /label="fusion site J5 (GTTA)"
     sig_peptide     728..787
                     /label="Ig-kappa signal peptide"
     region          788..1147
                     /label="trastuzumab VH"
     region          1148..2137
                     /label="human IgG1 constant CH1-hinge-CH2-CH3"
     sig_peptide     2731..2790
                     /label="Ig-kappa signal peptide"
     region          2791..3111
                     /label="trastuzumab VL"
     region          3112..3432
                     /label="human kappa constant"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctgtaagt atcaaggtta
      601 caagacaggt ttaaggagcc caatagaaac tgggcttgtc gagacagaga agactcttgc
      661 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacagagg
      721 agccaccatg gagacagaca ccctgctgct gtgggtgctg ctgctgtggg tgcccggctc
      781 tacaggcgag gtgcagctgg tggagtccgg aggaggactg gtgcaacccg gaggcagcct
      841 gcggttgtct tgcgctgcct ccggcttcaa catcaaggac acctacatcc actgggtgcg
      901 gcaggccccc ggcaaaggcc tggagtgggt tgctaggatc taccccacca acggctacac
      961 caggtacgcc gactctgtga agggccggtt caccatcagc gccgacacca gcaagaacac
     1021 cgcctacctg cagatgaaca gcctgcgggc cgaggacacc gccgtgtact actgttcccg
     1081 gtggggaggt gacggcttct acgccatgga ctactggggc cagggcaccc tggtgaccgt
     1141 gtcttctgcc tctaccaagg gcccctctgt gttccccctg gccccctctt ctaagtctac
     1201 ctccggcggc accgccgctt tgggctgcct ggtgaaggac tacttccccg agcccgtgac
     1261 cgtgtcttgg aacagcggcg ccctgacatc tggcgtgcac accttccccg ccgtgctgca
     1321 gtcttctggc ctgtacagcc tgtcctctgt ggtgaccgtg ccctcctctt ctctgggcac
     1381 ccagacctac atctgcaacg tgaaccacaa gccctccaac accaaggtgg acaagaaggt
     1441 ggagcccaag tcttgcgaca agacccacac ctgccctccc tgccccgccc ctgagttgtt
     1501 aggcggacct agcgtgttcc tgttcccccc taagcccaag gacaccctga tgatctccag
     1561 gacccccgag gtgacctgcg tggtggtgga cgtgtctcac gaggaccccg aggtgaagtt
     1621 caactggtac gtggacggcg tggaggtgca caacgccaag accaagcccc gggaggagca
     1681 gtacaactcc acctaccggg tggtgtctgt gctgaccgtg ctgcaccagg actggctgaa
     1741 cggcaaggag tacaagtgca aggtgtccaa caaggccctg cccgccccca tcgagaagac
     1801 catctccaag gccaagggcc agcctcggga gccccaggtg tacacattac ccccctcccg
     1861 ggatgagctg accaagaacc aggtgagcct gacctgcctg gtgaagggct tctacccctc
     1921 cgacatcgcc gtggagtggg agagcaacgg ccagcccgag aacaactaca agaccacacc
     1981 ccccgtgctg gacagcgacg gcagcttctt cctgtactcc aagctgaccg tggacaagag
     2041 ccggtggcag cagggcaacg tgttctcttg ctccgtgatg cacgaggccc tgcacaacca
     2101 ctacacccag aagtctctgt ccctgagccc cggcaagtaa tagacccctc tccctccccc
     2161 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     2221 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     2281 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     2341 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     2401 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     2461 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     2521 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     2581 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     2641 gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2701 gaaaaacacg atgataatat ggccacaacc atggaaaccg atacactcct cctctgggtc
     2761 ctcctcctct gggtccctgg aagcaccgga gacatccaga tgacccagag cccctcctct
     2821 ctgtctgcca gcgtgggcga ccgggtgacc atcacctgcc gggcttctca ggacgtgaac
     2881 accgccgtgg cctggtacca gcagaagccc ggcaaggccc ccaagttgct gatctactcc
     2941 gcctccttcc tgtactccgg cgtgccctct aggttcagcg gctctcggtc tggcaccgac
     3001 ttcaccctga ccatctcttc tctgcagccc gaggacttcg ccacctacta ctgccagcag
     3061 cactacacca cccctcccac cttcggccag ggcaccaagg tggagatcaa gcggaccgtg
     3121 gccgcccctt ctgtgttcat cttcccccct tctgacgagc agctgaagtc tggcaccgcc
     3181 tccgtggtgt gcctgctgaa caacttctac ccccgggagg ccaaggtgca gtggaaggtg
     3241 gacaacgccc tgcagtccgg caacagccag gagtccgtga ccgagcagga cagcaaggac
     3301 tccacctact ccctgagcag caccctgacc ctgtctaagg ccgactacga gaagcacaag
     3361 gtgtacgcct gcgaggtgac ccaccagggc ctgtcttctc ccgtgaccaa gtctttcaac
     3421 cggggcgagt gctaaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3481 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3541 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3601 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3661 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3721 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3781 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3841 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3901 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3961 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4021 ccgcctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4081 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4141 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4201 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg cgagtgtgtc
     4261 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4441 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4501 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4561 ttggaggcct aggcttttgc aaagccacca tgaccgagta caagcccacc gtgcggctgg
     4621 ccacccggga cgacgttcct agggctgtta ggaccctggc tgccgccttc gccgactacc
     4681 ccgccacaag gcacaccgtt gaccctgacc ggcacatcga gcgggtgacc gagctgcagg
     4741 agctgttcct gacccgggtg ggactggaca tcggcaaggt gtgggtggcc gacgacggag
     4801 ctgccgtggc cgtgtggaca acacctgagt ctgtggaggc cggcgccgtg ttcgccgaga
     4861 tcggccctag gatggctgag ctgtctggca gccggctggc tgcccaacag cagatggagg
     4921 gcctgctggc tcctcacagg cccaaggagc ccgcctggtt cttggccacc gtgggagtgt
     4981 ctcccgacca ccagggcaag ggcctgggat ctgccgtggt gctgcctgga gtggaggctg
     5041 ctgaaagggc cggagtgcct gccttcctgg agacatctgc cccccggaac ctgcctttct
     5101 acgagcggct gggcttcacc gtgaccgccg acgtggaggt tcctgaggga cctcggacct
     5161 ggtgcatgac ccggaagccc ggcgcctaaa acttgtttat tgcagcttat aatggttaca
     5221 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5281 gtggtttgtc caaactcatc aatgtatctt atgaatttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     5881 cgcagaaaaa aaggatctca acgcggaacc cctatttgtt tatttttcta aatacattca
     5941 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6001 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6061 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6121 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6181 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6241 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6301 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6361 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6421 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6481 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6541 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6601 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6661 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6721 ggttctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6781 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6841 ggtgcctcac tgattaagca ttggtaagtt a
//
