LOCUS       V041842                 6551 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041842
VERSION     V041842
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..2071
                     /label="P2RX4(NM_002560)"
                     /note="P2RX4(NM_002560)"
                     /gene="P2RX4"
     polyA_signal    2118..2342
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2388..2816
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2830..3159
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3226..4020
                     /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    4194..4315
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4364..4380)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4388..4404
                     /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(4412..4442)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4457..4478
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4766..5351)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5522..6382)
                     /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(6383..6487)
                     /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 gcgggctgct gcgccgcgct ggcggccttc ctgttcgagt acgacacgcc
      961 gcgcatcgtg ctcatccgca gccgcaaagt ggggctcatg aaccgcgccg tgcaactgct
     1021 catcctggcc tacgtcatcg ggtgggtgtt tgtgtgggaa aagggctacc aggaaactga
     1081 ctccgtggtc agctccgtta cgaccaaggt caagggcgtg gctgtgacca acacttctaa
     1141 acttggattc cggatctggg atgtggcgga ttatgtgata ccagctcagg aggaaaactc
     1201 cctcttcgtc atgaccaacg tgatcctcac catgaaccag acacagggcc tgtgccccga
     1261 gattccagat gcgaccactg tgtgtaaatc agatgccagc tgtactgccg gctctgccgg
     1321 cacccacagc aacggagtct caacaggcag gtgcgtagct ttcaacgggt ctgtcaagac
     1381 gtgtgaggtg gcggcctggt gcccggtgga ggatgacaca cacgtgccac aacctgcttt
     1441 tttaaaggct gcagaaaact tcactctttt ggttaagaac aacatctggt atcccaaatt
     1501 taatttcagc aagaggaata tccttcccaa catcaccact acttacctca agtcgtgcat
     1561 ttatgatgct aaaacagatc ccttctgccc catattccgt cttggcaaaa tagtggagaa
     1621 cgcaggacac agtttccagg acatggccgt ggagggaggc atcatgggca tccaggtcaa
     1681 ctgggactgc aacctggaca gagccgcctc cctctgcttg cccaggtact ccttccgccg
     1741 cctcgataca cgggacgttg agcacaacgt atctcctggc tacaatttca ggtttgccaa
     1801 gtactacaga gacctggctg gcaacgagca gcgcacgctc atcaaggcct atggcatccg
     1861 cttcgacatc attgtgtttg ggaaggcagg gaaatttgac atcatcccca ctatgatcaa
     1921 catcggctct ggcctggcac tgctaggcat ggcgaccgtg ctgtgtgaca tcatagtcct
     1981 ctactgcatg aagaaaagac tctactatcg ggagaagaaa tataaatatg tggaagatta
     2041 cgagcagggt cttgctagtg agctggacca gtgactcgag tctagagggc ccgtttaaac
     2101 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2161 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2221 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2281 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2341 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2401 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2461 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2521 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2581 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2641 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2701 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2761 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2821 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2881 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3001 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3061 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3121 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3181 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3241 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3301 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3361 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3421 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3481 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3541 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3601 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3661 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3721 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3781 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3841 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3901 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3961 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4021 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4081 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4141 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4321 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4381 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4441 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4501 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4561 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4621 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4681 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4741 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4801 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4861 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4921 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4981 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5041 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5101 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5161 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5221 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5281 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5341 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5401 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5461 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5521 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5641 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5821 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6361 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6481 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6541 tcccctatgg t
//
