LOCUS       sequence-433366-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses anti-HER2 39S Light chain in mammalian cells. To be paired
            with 39S heavy chain to form the anti-HER2 39S Fab.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4630)
  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 4630)
  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: pcDNA3.1-39S Light chain
FEATURES             Location/Qualifiers
     source          1..4630
                     /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(1906..1923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1912..2136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2182..2610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2269..2288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2479..2500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2619..2639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2624..2953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2804..2939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2866..2885
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3020..3814
                     /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(3074..3093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3684..3703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3988..4109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4025..4044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4079..4098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4158..4174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4158..4174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4171..4193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4182..4198
                     /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(4206..4212,4213..4230,4231..4236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4251..4272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4389..4406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(4495..4599)
                     /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 gatgtgtagc
     1021 tgacattgtg atgacgcaaa cccctttatc tctgtccgtc acgccggggc agccagcttc
     1081 aatttcttgc aagtcgtcac agtccgtgtt tttccgctcg aacaataaaa atatcttagc
     1141 ttggtatttg cagaagcctg gccaaccccc gcaattactg atctactggg cgagttcacg
     1201 cgaatcaggc gtaccggacc gtttttcagg atcggggagc ggcaccgatt ttacactgaa
     1261 gatttctcgt gtggaggcgg aagacgtcgg agtttactat tgccaacaat atttcggttc
     1321 cccattcacc tttgggccag gtaccaaagt cgatattaag aggactgtgg ctgcaccatc
     1381 tgtcttcatc ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg
     1441 cctgctgaat aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct
     1501 ccaatcgggt aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag
     1561 cctcagcagc accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg
     1621 cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg
     1681 ttgataaacc ggtaagctga gcagcggcta caaggacgtg atcctgtggt tcagcttcgg
     1741 cgccagctgc ttcatcctgc tggccatcgt gatgggcctg gtgttcatct gcgtgaagaa
     1801 cggcaacatg cggtgcacca tctgcatctg atgatatcga attccaccac actggactag
     1861 tggatccgag ctcggtacca agcttaagtt taaaccgctg atcagcctcg actgtgcctt
     1921 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1981 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2041 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2101 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2161 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2281 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2341 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2401 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2461 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2521 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2581 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     2641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2701 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2881 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2941 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3001 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3181 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3541 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3661 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3781 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3841 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3901 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3961 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4081 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4141 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4201 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4261 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaat
     4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4621 acctgacgtc
//
