LOCUS       V038073                 7069 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038073
VERSION     V038073
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2505
                     /label="LAG3(NM_002286)"
                     /note="LAG3(NM_002286)"
                     /gene="LAG3"
     CDS             2512..2601
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2636..2860
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2906..3334
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3348..3677
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3744..4538
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4712..4833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4882..4898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4906..4922
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4930..4960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4975..4996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5284..5869)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6040..6900)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtgggagg ctcagttcct gggcttgctg
      961 tttctgcagc cgctttgggt ggctccagtg aagcctctcc agccaggggc tgaggtcccg
     1021 gtggtgtggg cccaggaggg ggctcctgcc cagctcccct gcagccccac aatccccctc
     1081 caggatctca gccttctgcg aagagcaggg gtcacttggc agcatcagcc agacagtggc
     1141 ccgcccgctg ccgcccccgg ccatcccctg gcccccggcc ctcacccggc ggcgccctcc
     1201 tcctgggggc ccaggccccg ccgctacacg gtgctgagcg tgggtcccgg aggcctgcgc
     1261 agcgggaggc tgcccctgca gccccgcgtc cagctggatg agcgcggccg gcagcgcggg
     1321 gacttctcgc tatggctgcg cccagcccgg cgcgcggacg ccggcgagta ccgcgccgcg
     1381 gtgcacctca gggaccgcgc cctctcctgc cgcctccgtc tgcgcctggg ccaggcctcg
     1441 atgactgcca gccccccagg atctctcaga gcctccgact gggtcatttt gaactgctcc
     1501 ttcagccgcc ctgaccgccc agcctctgtg cattggttcc ggaaccgggg ccagggccga
     1561 gtccctgtcc gggagtcccc ccatcaccac ttagcggaaa gcttcctctt cctgccccaa
     1621 gtcagcccca tggactctgg gccctggggc tgcatcctca cctacagaga tggcttcaac
     1681 gtctccatca tgtataacct cactgttctg ggtctggagc ccccaactcc cttgacagtg
     1741 tacgctggag caggttccag ggtggggctg ccctgccgcc tgcctgctgg tgtggggacc
     1801 cggtctttcc tcactgccaa gtggactcct cctgggggag gccctgacct cctggtgact
     1861 ggagacaatg gcgactttac ccttcgacta gaggatgtga gccaggccca ggctgggacc
     1921 tacacctgcc atatccatct gcaggaacag cagctcaatg ccactgtcac attggcaatc
     1981 atcacagtga ctcccaaatc ctttgggtca cctggatccc tggggaagct gctttgtgag
     2041 gtgactccag tatctggaca agaacgcttt gtgtggagct ctctggacac cccatcccag
     2101 aggagtttct caggaccttg gctggaggca caggaggccc agctcctttc ccagccttgg
     2161 caatgccagc tgtaccaggg ggagaggctt cttggagcag cagtgtactt cacagagctg
     2221 tctagcccag gtgcccaacg ctctgggaga gccccaggtg ccctcccagc aggccacctc
     2281 ctgctgtttc tcatccttgg tgtcctttct ctgctccttt tggtgactgg agcctttggc
     2341 tttcaccttt ggagaagaca gtggcgacca agacgatttt ctgccttaga gcaagggatt
     2401 caccctccgc aggctcagag caagatagag gagctggagc aagaaccgga gccggagccg
     2461 gagccggaac cggagcccga gcccgagccc gagccggagc agctcctcga ggaacaaaaa
     2521 ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt agagcaaaag
     2581 ttaatttccg aggaggactt atgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2641 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2701 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2761 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2821 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2881 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3301 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3601 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3661 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3721 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3781 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3841 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3901 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3961 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4021 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4081 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4141 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4201 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4261 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4321 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4381 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4441 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4501 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4561 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4621 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4681 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4801 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4861 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5701 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
