LOCUS       sequence-75289-e        8574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human IgG1/kappa antibody isotype with unknown 
            specificity. Useful vector for cloning of human antibodies with 
            kappa light chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8574)
  AUTHORS   Dodev TS, Karagiannis P, Gilbert AE, Josephs DH, Bowen H, James LK, 
            Bax HJ, Beavil R, Pang MO, Gould HJ, Karagiannis SN, Beavil AJ
  TITLE     A tool kit for rapid cloning and expression of recombinant 
            antibodies.
  JOURNAL   Sci Rep. 2014 Jul 30;4:5885. doi: 10.1038/srep05885.
  PUBMED    25073855
REFERENCE   2  (bases 1 to 8574)
  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: "Sci Rep."; 
            date: "2014-07-30"; volume: "4:5885. doi"; pages: " 
            10.1038/srep05885"
COMMENT     Sequence Label: pVITRO1-M80-F2-IgG1/kappa
FEATURES             Location/Qualifiers
     source          1..8574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    3006..3127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3071..3090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3315..3903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3804..3823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3983..4286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4401..5706
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    4743..5674
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     misc_feature    6421..6865
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6898..6944
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6936..6954
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6964..7989
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    8001..8573
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtatg gactggacct ggaggatcct cttcttggtg gcggccgcca caggcgcgca
     1621 ctccgaggtg cagctggtgg agtctggggg aggcctggta cagcctgggg ggtccctgag
     1681 actctcctgt gcagcctctg gattcacctt taggcactat gccatcagtt gggtccgcca
     1741 ggctccaggg aaggggctgg agtgggtctc aggtctgagt ggtagtggaa ataggacata
     1801 ctacgcagac tccgtgaagg gccggttcac catctccaga gacaattccg agaacacgct
     1861 gtttttgcaa atgaacagcc tgagagccga ggacacggcc gtgtattact gtgcgaaaga
     1921 tcgccgagtg ggagctacct tcgtctttga ctcctggggc cagggaaccc tggtcaccgt
     1981 ctccccagct agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac
     2041 ctctgggggc acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac
     2101 ggtgtcgtgg aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca
     2161 gtcctcagga ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac
     2221 ccagacctac atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt
     2281 tgagcccaaa tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct
     2341 ggggggaccg tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg
     2401 gacccctgag gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt
     2461 caactggtac gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca
     2521 gtacaacagc acgtaccggg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa
     2581 tggcaaggag tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac
     2641 catctccaaa gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg
     2701 ggatgagctg accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag
     2761 cgacatcgcc gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc
     2821 tcccgtgctg gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag
     2881 caggtggcag caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca
     2941 ctacacgcag aagagcctct ccctgtctcc gggtaaatga tgtacagcta gctggccaga
     3001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3181 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaaatgttaa ttaactagcc
     3241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3421 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3961 atgttcttaa ttaacctgca ggcgttacat aacttacggt aaatggcccg cctggctgac
     4021 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4081 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4141 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4201 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4261 acgtattagt catcgctatt accatgatga tgcggttttg gcagtacatc aatgggcgtg
     4321 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4381 tgttttgact agtggagccg agagtaattc atacaaaagg agggatcgcc ttcgcaaggg
     4441 gagagcccag ggaccgtccc taaattctca cagacccaaa tccctgtagc cgccccacga
     4501 cagcgcgagg agcatgcgcc cagggctgag cgcgggtaga tcagagcaca caagctcaca
     4561 gtccccggcg gtggggggag gggcgcgctg agcgggggcc agggagctgg cgcggggcaa
     4621 actgggaaag tggtgtcgtg tgctggctcc gccctcttcc cgagggtggg ggagaacggt
     4681 atataagtgc ggtagtcgcc ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc
     4741 aggtgagtgg cgggtgtggc ttccgcgggc cccggagctg gagccctgct ctgagcgggc
     4801 cgggctgata tgcgagtgtc gtccgcaggg tttagctgtg agcattccca cttcgagtgg
     4861 cgggcggtgc gggggtgaga gtgcgaggcc tagcggcaac cccgtagcct cgcctcgtgt
     4921 ccggcttgag gcctagcgtg gtgtccgccg ccgcgtgcca ctccggccgc actatgcgtt
     4981 ttttgtcctt gctgccctcg attgccttcc agcagcatgg gctaacaaag ggagggtgtg
     5041 gggctcactc ttaaggagcc catgaagctt acgttggata ggaatggaag ggcaggaggg
     5101 gcgactgggg cccgcccgcc ttcggagcac atgtccgacg ccacctggat ggggcgaggc
     5161 ctgtggcttt ccgaagcaat cgggcgtgag tttagcctac ctgggccatg tggccctagc
     5221 actgggcacg gtctggcctg gcggtgccgc gttcccttgc ctcccaacaa gggtgaggcc
     5281 gtcccgcccg gcaccagttg cttgcgcgga aagatggccg ctcccggggc cctgttgcaa
     5341 ggagctcaaa atggaggacg cggcagcccg gtggagcggg cgggtgagtc acccacacaa
     5401 aggaagaggg ccttgcccct cgccggccgc tgcttcctgt gaccccgtgg tctatcggcc
     5461 gcatagtcac ctcgggcttc tcttgagcac cgctcgtcgc ggcgggggga ggggatctaa
     5521 tggcgttgga gtttgttcac atttggtggg tggagactag tcaggccagc ctggcgctgg
     5581 aagtcattct tggaatttgc ccctttgagt ttggagcgag gctaattctc aagcctctta
     5641 gcggttcaaa ggtattttct aaacccgttt ccaggtgttg tgaaagccac cgctaattca
     5701 aagcaatccg gaatgttgcc atcacaactc attgggtttc tgctgctctg ggttccagct
     5761 agccgcggtg acatccagat gacccagtct ccatccaccc tgtctgcatc tgtgggagac
     5821 agagtcacca tcacttgccg ggcaagtcag aacattacca cctatttcaa ttggtatcag
     5881 caaaaaccag ggaaagcccc taaactcctg atctatgctg catccagttt gcaaagtggg
     5941 gtcccatcaa ggttcagtgg cagtggatct gggacagatt tcactctcac catcaccagt
     6001 ctgcaacctg aagattttgc aacttactac tgtcaacaga gggggacgtt cggccaaggg
     6061 accaaggtgg aaatcaaacg tacggtggcg gcgccatctg tcttcatctt cccgccatct
     6121 gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc
     6181 agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag
     6241 agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg
     6301 agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg
     6361 agctcgcccg tcacaaagag cttcaacagg ggagagtgtt agggatcccg tacgcctagg
     6421 agcaggtttc cccaatgaca caaaacgtgc aacttgaaac tccgcctggt ctttccaggt
     6481 ctagaggggt aacactttgt actgcgtttg gctccacgct cgatccactg gcgagtgtta
     6541 gtaacagcac tgttgcttcg tagcggagca tgacggccgt gggaactcct ccttggtaac
     6601 aaggacccac ggggccaaaa gccacgccca cacgggcccg tcatgtgtgc aaccccagca
     6661 cggcgacttt actgcgaaac ccactttaaa gtgacattga aactggtacc cacacactgg
     6721 tgacaggcta aggatgccct tcaggtaccc cgaggtaaca cgcgacactc gggatctgag
     6781 aaggggactg gggcttctat aaaagcgctc ggtttaaaaa gcttctatgc ctgaataggt
     6841 gaccggaggt cggcaccttt cctttgcaat tactgaccct atgaatacac tgactgtttg
     6901 acaattaatc atcggcatag tatatcggca tagtataata cgactcacta taggagggcc
     6961 accatgaaga aacctgaact gacagcaact tctgttgaga agtttctcat tgaaaaattt
     7021 gattctgttt ctgatctcat gcagctgtct gaaggtgaag aaagcagagc cttttctttt
     7081 gatgttggag gaagaggtta tgttctgagg gtcaattctt gtgctgatgg tttttacaaa
     7141 gacagatatg tttacagaca ctttgcctct gctgctctgc caattccaga agttctggac
     7201 attggagaat tttctgaatc tctcacctac tgcatcagca gaagagcaca aggagtcact
     7261 ctccaggatc tccctgaaac tgagctgcca gctgttctgc aacctgttgc tgaagcaatg
     7321 gatgccattg cagcagctga tctgagccaa acctctggat ttggtccttt tggtccccaa
     7381 ggcattggtc agtacaccac ttggagggat ttcatttgtg ccattgctga tcctcatgtc
     7441 tatcactggc agactgtgat ggatgacaca gtttctgctt ctgttgctca ggcactggat
     7501 gaactcatgc tgtgggcaga agattgtcct gaagtcagac acctggtcca tgctgatttt
     7561 ggaagcaaca atgttctgac agacaatggc agaatcactg cagtcattga ctggtctgaa
     7621 gccatgtttg gagattctca atatgaggtt gccaacattt ttttttggag accttggctg
     7681 gcttgcatgg aacaacaaac aagatatttt gaaagaagac acccagaact ggctggttcc
     7741 cccagactga gagcctacat gctcagaatt ggcctggacc aactgtatca atctctggtt
     7801 gatggaaact ttgatgatgc tgcttgggca caaggaagat gtgatgccat tgtgaggtct
     7861 ggtgctggaa ctgttggaag aactcaaatt gcaagaaggt ctgctgctgt ttggactgat
     7921 ggatgtgttg aagttctggc tgactctgga aacaggagac cctccacaag acccagagcc
     7981 aaggaatgaa tattagctag attatcccta atacctgcca ccccactctt aatcagtggt
     8041 ggaagaacgg tctcagaact gtttgtttca attggccatt taagtttagt agtaaaagac
     8101 tggttaatga taacaatgca tcgtaaaacc ttcagaagga aaggagaatg ttttgtggac
     8161 cactttggtt ttcttttttg cgtgtggcag ttttaagtta ttagttttta aaatcagtac
     8221 tttttaatgg aaacaacttg accaaaaatt tgtcacagaa ttttgagacc cattaaaaaa
     8281 gttaaatgag aaacctgtgt gttcctttgg tcaacaccga gacatttagg tgaaagacat
     8341 ctaattctgg ttttacgaat ctggaaactt cttgaaaatg taattcttga gttaacactt
     8401 ctgggtggag aatagggttg ttttcccccc acataattgg aaggggaagg aatatcattt
     8461 aaagctatgg gagggttgct ttgattacaa cactggagag aaatgcagca tgttgctgat
     8521 tgcctgtcac taaaacaggc caaaaactga gtccttgggt tgcatagaaa gctg
//
