LOCUS       V040212                 7671 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040212
VERSION     V040212
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             908..2482
                     /label="LAG3(NM_002286)"
                     /note="LAG3(NM_002286)"
                     /gene="LAG3"
     CDS             2531..3187
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK*"
                     /label="GST"
     polyA_signal    3238..3462
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3508..3936
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3950..4279
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4346..5140
                     /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    5314..5435
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5484..5500)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5508..5524
                     /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(5532..5562)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5577..5598
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5886..6471)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6642..7502)
                     /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(7503..7607)
                     /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 cgccaccatg tgggaggctc agttcctggg cttgctgttt ctgcagccgc tttgggtggc
      961 tccagtgaag cctctccagc caggggctga ggtcccggtg gtgtgggccc aggagggggc
     1021 tcctgcccag ctcccctgca gccccacaat ccccctccag gatctcagcc ttctgcgaag
     1081 agcaggggtc acttggcagc atcagccaga cagtggcccg cccgctgccg cccccggcca
     1141 tcccctggcc cccggccctc acccggcggc gccctcctcc tgggggccca ggccccgccg
     1201 ctacacggtg ctgagcgtgg gtcccggagg cctgcgcagc gggaggctgc ccctgcagcc
     1261 ccgcgtccag ctggatgagc gcggccggca gcgcggggac ttctcgctat ggctgcgccc
     1321 agcccggcgc gcggacgccg gcgagtaccg cgccgcggtg cacctcaggg accgcgccct
     1381 ctcctgccgc ctccgtctgc gcctgggcca ggcctcgatg actgccagcc ccccaggatc
     1441 tctcagagcc tccgactggg tcattttgaa ctgctccttc agccgccctg accgcccagc
     1501 ctctgtgcat tggttccgga accggggcca gggccgagtc cctgtccggg agtcccccca
     1561 tcaccactta gcggaaagct tcctcttcct gccccaagtc agccccatgg actctgggcc
     1621 ctggggctgc atcctcacct acagagatgg cttcaacgtc tccatcatgt ataacctcac
     1681 tgttctgggt ctggagcccc caactccctt gacagtgtac gctggagcag gttccagggt
     1741 ggggctgccc tgccgcctgc ctgctggtgt ggggacccgg tctttcctca ctgccaagtg
     1801 gactcctcct gggggaggcc ctgacctcct ggtgactgga gacaatggcg actttaccct
     1861 tcgactagag gatgtgagcc aggcccaggc tgggacctac acctgccata tccatctgca
     1921 ggaacagcag ctcaatgcca ctgtcacatt ggcaatcatc acagtgactc ccaaatcctt
     1981 tgggtcacct ggatccctgg ggaagctgct ttgtgaggtg actccagtat ctggacaaga
     2041 acgctttgtg tggagctctc tggacacccc atcccagagg agtttctcag gaccttggct
     2101 ggaggcacag gaggcccagc tcctttccca gccttggcaa tgccagctgt accaggggga
     2161 gaggcttctt ggagcagcag tgtacttcac agagctgtct agcccaggtg cccaacgctc
     2221 tgggagagcc ccaggtgccc tcccagcagg ccacctcctg ctgtttctca tccttggtgt
     2281 cctttctctg ctccttttgg tgactggagc ctttggcttt cacctttgga gaagacagtg
     2341 gcgaccaaga cgattttctg ccttagagca agggattcac cctccgcagg ctcagagcaa
     2401 gatagaggag ctggagcaag aaccggagcc ggagccggag ccggaaccgg agcccgagcc
     2461 cgagcccgag ccggagcagc tcggtggagg cggatcagga ggcggtggat ctccgaattc
     2521 tgcagatatc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     2581 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     2641 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     2701 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     2761 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     2821 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     2881 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     2941 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     3001 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     3061 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     3121 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     3181 aaaatgagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3241 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3301 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3361 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3421 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3481 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3721 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3841 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4321 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4381 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4441 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4501 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4561 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4621 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4681 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4741 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4801 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4861 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4921 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4981 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5041 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5101 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5161 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5221 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5281 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5341 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5401 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5461 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5521 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5581 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5641 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5701 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5821 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5881 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5941 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6001 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6061 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6121 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6181 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6241 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6301 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6361 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6421 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6481 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6541 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6601 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6661 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6721 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6781 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6841 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6901 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6961 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7021 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7081 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7141 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7201 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7261 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7321 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7381 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7441 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7501 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7561 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7621 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
