LOCUS       sequence-181655-        5741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of secretory immunoglobulin heavy chains in mammalian 
            cells, human IgG1 isotype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  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 5741)
  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: AbVec2.0-IGHG1
FEATURES             Location/Qualifiers
     source          1..5741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    298..319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(334..339,340..357,358..364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    372..388
                     /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     377..399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     396..412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     396..412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        441..820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        821..1024
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     975..995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1021..1045
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1255..1273
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1255..1272
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1286..1305
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2468..2477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2481..2615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2518..2537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2572..2591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2684..3041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2684..2875
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2707..2727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2892..3027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3061..3078)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3061..3077)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3070..3092)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3290..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3377..3396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3587..3608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3762..3781)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3881..3903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3941..3959)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4027..4131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4132..4200,4201..4992)
                     /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(4350..4369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5163..5741,1..10)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5652..5671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
       61 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      181 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      241 atccaactgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      301 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      361 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac
      421 gaattaattc gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac
      481 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      541 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      601 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      661 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      721 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      781 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      841 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      901 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      961 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1021 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     1081 tagaagacac cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat
     1141 tccccgtgcc aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc
     1201 ttcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     1261 gtgacactat agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc
     1321 caactgcacc tcggttctat cgattgaatt ccaccatggg atggtcatgt atcatccttt
     1381 ttctagtagc aactgcaacc ggttgactaa ctagctggtc gaccaagggc ccaagcgtct
     1441 tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg ggctgcctgg
     1501 tcaaggacta cttccccgag ccggtgacgg tgtcgtggaa ctcaggcgcc ctgaccagcg
     1561 gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc agcagcgtgg
     1621 tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg aatcacaagc
     1681 ccagcaacac caaggtggac aagagagttg agcccaaatc ttgtgacaaa actcacacat
     1741 gcccaccgtg cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa
     1801 aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg
     1861 tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata
     1921 atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc
     1981 tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca
     2041 aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac
     2101 cacaggtgta caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga
     2161 cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc
     2221 agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc
     2281 tctatagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct
     2341 ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtccccgg
     2401 gtaaatgagt gcgacggccg gcaagccccc gctccccggg ctctcggggt cgcgcgagga
     2461 aagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca
     2521 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2581 ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat
     2641 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
     2701 atctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2761 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3001 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc
     3061 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3121 ccttgcagca catccccctt tcgccagttg gcgtaatagc gaagaggccc gcaccgatcg
     3181 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     3241 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3481 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3601 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3721 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3781 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3901 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3961 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4021 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4081 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4141 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4201 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4261 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4321 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4381 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4441 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4501 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4561 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4621 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4681 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4741 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4801 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4861 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4921 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4981 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5041 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5101 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5281 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5341 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5401 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5461 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5521 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5581 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5641 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5701 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga g
//
