LOCUS       sequence-182241-        4976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of immunoglobulin light chains in mammalian cells, human 
            lambda 2 constant region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  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-IGLC2-XhoI
FEATURES             Location/Qualifiers
     source          1..4976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        249..628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        629..832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     783..803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     829..853
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1063..1081
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1063..1080
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1094..1113
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1511..1520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1524..1658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1561..1580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1615..1634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1727..2084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1727..1918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1750..1770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1935..2070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2104..2121)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2104..2120)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2113..2135)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2333..2788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2420..2439)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2630..2651
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2805..2824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2924..2946
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2984..3002)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3070..3174
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3175..3243,3244..4035)
                     /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(3393..3412)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4206..4794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4695..4714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4948..4965
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat ccaactggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga attaattcga
      241 gctcgcccga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      841 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      901 ggaccgatcc agcctccgcg gccgggaacg gtgcattgga acgcggattc cccgtgccaa
      961 gagtgacgta agtaccgcct atagagtcta taggcccacc cccttggctt cgttagaacg
     1021 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
     1081 aataacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca actgcacctc
     1141 ggttctatcg attgaattcc accatgggat ggtcatgtat catccttttt ctagtagcaa
     1201 ctgcaaccgg ttgactaact agcctcgagt gaggagcttc aagccaacaa ggccacactg
     1261 gtgtgtctca taagtgactt ctacccggga gccgtgacag tggcctggaa ggcagatagc
     1321 agccccgtca aggcgggagt ggagaccacc acaccctcca aacaaagcaa caacaagtac
     1381 gcggccagca gctacctgag cctgacgcct gagcagtgga agtcccacag aagctacagc
     1441 tgccaggtca cgcatgaagg gagcaccgtg gagaagacag tggcccctac agaatgttca
     1501 tagaagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa
     1561 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1621 tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac
     1681 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     1741 accatctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt
     2101 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     2161 tcgccttgca gcacatcccc ctttcgccag ttggcgtaat agcgaagagg cccgcaccga
     2221 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     2281 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2461 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2521 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2581 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2641 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2701 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2761 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2821 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2881 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2941 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3001 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3061 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3121 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3181 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3241 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3301 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3361 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3421 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3481 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3541 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3601 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3661 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3721 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3781 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3841 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3901 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3961 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4021 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4081 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4141 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4201 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4261 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4321 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4381 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4441 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4501 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4561 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4621 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4681 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4741 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4801 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4861 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4921 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
