LOCUS       sequence-181656-        5018 bp    DNA     circular SYN 14-APR-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 5018)
  AUTHORS   Tiller T, Busse CE, Wardemann H
  TITLE     Cloning and expression of murine Ig genes from single B cells.
  JOURNAL   J Immunol Methods. 2009 Oct 31;350(1-2):183-93. doi: 
            10.1016/j.jim.2009.08.009. Epub 2009 Aug 27.
  PUBMED    19716372
REFERENCE   2  (bases 1 to 5018)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Immunol 
            Methods."; date: "2009-10-31"; volume: "350(1-2):183-93. doi"; 
            pages: " 10.1016/j.jim.2009.08.009. Epub 2009 Aug 27"
COMMENT     Sequence Label: AbVec2.1-IGLC2-MscI
FEATURES             Location/Qualifiers
     source          1..5018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    51..72
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(87..92,93..110,111..117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    125..141
                     /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     130..152
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     149..165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     149..165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        194..573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        574..777
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     728..748
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     774..798
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1008..1026
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1008..1025
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1039..1058
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1498..1507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1511..1645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1548..1567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1602..1621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1714..2071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1714..1905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1737..1757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1922..2057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2091..2108)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2091..2107)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2100..2122)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2320..2775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2407..2426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2617..2638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2792..2811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2911..2933
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2971..2989)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3057..3161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3162..3230,3231..4022)
                     /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(3380..3399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4193..4781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4682..4701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4935..4952
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
       61 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      121 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattaa
      181 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
      241 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      301 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      361 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      421 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      481 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      541 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      601 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      661 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      721 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      781 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      841 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      901 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      961 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
     1021 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
     1081 acctcggttc tatcgattga attccaccat gggatggtca tgtatcatcc tttttctagt
     1141 agcaactgca accggttgac taactagctg gccagcccaa ggctgccccc tcggtcactc
     1201 tgttcccgcc ttcgagtgag gagcttcaag ccaacaaggc cacactggtg tgtctcataa
     1261 gtgacttcta cccgggagcc gtgacagtgg cctggaaggc agatagcagc cccgtcaagg
     1321 cgggagtgga gaccaccaca ccctccaaac aaagcaacaa caagtacgcg gccagcagct
     1381 acctgagcct gacgcctgag cagtggaagt cccacagaag ctacagctgc caggtcacgc
     1441 atgaagggag caccgtggag aagacagtgg cccctacaga atgttcatag aagcttggcc
     1501 gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1621 caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat
     1681 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc atctgtggaa
     1741 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     1801 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     1921 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1981 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2041 aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc
     2101 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2161 catccccctt tcgccagttg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2221 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2281 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2521 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2641 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2701 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2761 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2821 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2881 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2941 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3001 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3061 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3181 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3241 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3301 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3361 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3421 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3481 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3541 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3601 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3661 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3721 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3781 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3841 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3901 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3961 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4021 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4081 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4141 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4201 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4261 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4321 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4381 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4441 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4501 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4561 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4621 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4681 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4741 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4801 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4861 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4921 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4981 cgcctctccc cgcgcgttgg ccgattcatt aatccaac
//
