LOCUS       sequence-443901-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses anti-HER2 Tras Light chain in mammalian cells. To be 
            paired with Tras heavy chain to form the anti-HER2 Tras Fab.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   Driscoll CL, Keeble AH, Howarth MR
  TITLE     SpyMask enables combinatorial assembly of bispecific binders.
  JOURNAL   Nat Commun. 2024 Mar 16;15(1):2403. doi: 10.1038/s41467-024-46599-9.
  PUBMED    38493197
REFERENCE   2  (bases 1 to 6306)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2024-03-16"; volume: "15(1):2403. doi"; pages: " 
            10.1038/s41467-024-46599-9"
COMMENT     Sequence Label: pcDNA3.1-Tras light chain
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      932..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1897..1914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1903..2127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2173..2601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2260..2279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2470..2491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2610..2630)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2615..2944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2795..2930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2857..2876
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3011..3805
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3065..3084)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3675..3694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3979..4100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4016..4035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4070..4089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4149..4165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4149..4165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4162..4184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4173..4189
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4197..4203,4204..4221,4222..4227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4242..4263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4380..4397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4551..5139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4631..4650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5310..6101,6102..6170))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5933..5952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6171..6275)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccaccatg gggatccttc ccagccctgg
      961 gatgcctgcg ctgctctccc tcgtgagcct tctctccgtg ctgctgatgg gttgcgtagc
     1021 tggtaccgga gacatccaga tgacccagag tccctcttcc ctgagcgcca gcgtggggga
     1081 cagggtgacc ataacttgca gggcgtctca agacgtgaac accgccgtgg cctggtatca
     1141 acagaagccg ggcaaagccc ccaaactgct gatctatagc gcctccttcc tctacagcgg
     1201 cgtgccctct aggttcagtg gtagccgcag cggcaccgac tttaccctta ctatcagctc
     1261 cctccagccc gaagacttcg ccacctacta ctgccaacag cactacacca ccccccctac
     1321 ctttggccag ggcaccaaag tggagatcaa gaggaccgtg gctgccccaa gcgtgttcat
     1381 cttccccccc agcgacgagc agctgaagag cggtacggct agcgtggtgt gcctgctgaa
     1441 caacttctac cctcgagagg ccaaggtgca gtggaaggtg gacaatgcgt tgcagagcgg
     1501 caactcccag gagagcgtga ccgagcagga cagcaaagac agcacctaca gccttagctc
     1561 aacactcacc ttgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac
     1621 ccatcagggt ctgagtagtc ccgtgactaa gagcttcaac aggggcgagt gctgataaac
     1681 cggtaagctg agcagcggct acaaggacgt gatcctgtgg ttcagcttcg gcgccagctg
     1741 cttcatcctg ctggccatcg tgatgggcct ggtgttcatc tgcgtgaaga acggcaacat
     1801 gcggtgcacc atctgcatct gatgatatcg aattccacca cactggacta gtggatccga
     1861 gctcggtacc aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     1921 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1981 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2041 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2101 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2161 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2221 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2281 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2341 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2401 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2461 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2521 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2581 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2641 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2701 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2761 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2821 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2881 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2941 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3001 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3061 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3121 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3181 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3241 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3301 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3361 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3421 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3481 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3541 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3601 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3661 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3721 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3781 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3841 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3901 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3961 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4021 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4081 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4141 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4201 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4261 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4321 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4381 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4981 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5101 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5221 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5281 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5341 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5401 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5461 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5521 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5581 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5641 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5701 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5761 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5821 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5881 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5941 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6001 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6061 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6121 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6181 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6241 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6301 gacgtc
//
