LOCUS       sequence-181657-        5020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of immunoglobulin light chains in mammalian cells, human 
            kappa constant region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5020)
  AUTHORS   Tiller T, Meffre E, Yurasov S, Tsuiji M, Nussenzweig MC, Wardemann H
  TITLE     Efficient generation of monoclonal antibodies from single human B 
            cells by single cell RT-PCR and expression vector cloning.
  JOURNAL   J Immunol Methods. 2008 Jan 1;329(1-2):112-24. Epub 2007 Oct 31.
  PUBMED    17996249
REFERENCE   2  (bases 1 to 5020)
  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: "J Immunol 
            Methods."; date: "2008-01-1"; volume: "329(1-2)"; pages: "112-24. 
            Epub 2007 Oct 31"
COMMENT     Sequence Label: AbVec1.1-IGKC
FEATURES             Location/Qualifiers
     source          1..5020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    92..113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(128..133,134..151,152..158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    166..182
                     /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)."
     primer_bind     171..193
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     190..206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     190..206
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     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     815..839
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1049..1067
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1049..1066
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1080..1099
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1541..1550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1554..1688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1591..1610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1645..1664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1757..2114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1757..1948
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1780..1800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1965..2100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2134..2151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2134..2150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2363..2818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2450..2469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2660..2681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2835..2854)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2954..2976
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3014..3032)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3100..3204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3205..3273,3274..4065)
                     /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     complement(3423..3442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4236..4824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4725..4744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4978..4995
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatccaa ctggcacgac aggtttcccg
       61 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      121 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      181 aatttcacac aggaaacagc tatgacatga ttacgaatta attcgagctc gcccgacatt
      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 gcagagctcg tttagtgaac cgtcagatcg
      841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta
      961 ccgcctatag agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat
     1021 acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt
     1081 tgcctttctc tccacaggtg tccactccca ggtccaactg cacctcggtt ctatcgattg
     1141 aattccacca tgggatggtc atgtatcatc ctttttctag tagcaactgc aaccggttga
     1201 ctaactagcc gtacggtggc tgcaccatct gtcttcatct tcccgccatc tgatgagcag
     1261 ttgaaatctg gaactgcctc tgttgtgtgc ctgctgaata acttctatcc cagagaggcc
     1321 aaagtacagt ggaaggtgga taacgccctc caatcgggta actcccagga gagtgtcaca
     1381 gagcaggaca gcaaggacag cacctacagc ctcagcagca ccctgacgct gagcaaagca
     1441 gactacgaga aacacaaagt ctacgcctgc gaagtcaccc atcagggcct gagctcgccc
     1501 gtcacaaaga gcttcaacag gggagagtgt tagaagcttg gccgccatgg cccaacttgt
     1561 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1621 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1681 tctggatcgg gaattaattc ggcgcagcac catggcctga aataacctct gaaagaggaa
     1741 cttggttagg taccttctga ggcggaaaga accatctgtg gaatgtgtgt cagttagggt
     1801 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1861 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1921 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1981 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2041 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2101 taggcttttg caaaaagctg ttaacagctt ggcactggcc gtcgttttac aacgtcgtga
     2161 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2221 ttggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2281 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2341 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2401 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2461 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2521 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2581 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2641 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2701 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2761 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     2821 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2881 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2941 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3001 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3061 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
//
