LOCUS       V039847                 6611 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039847
VERSION     V039847
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"
     regulatory      902..911
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             911..934
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1232..2131
                     /label="OPRM1(NM_001285526)"
                     /note="OPRM1(NM_001285526)"
                     /gene="OPRM1"
     polyA_signal    2178..2402
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2448..2876
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2890..3219
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3286..4080
                     /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    4254..4375
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4424..4440)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4448..4464
                     /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(4472..4502)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4517..4538
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4826..5411)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5582..6442)
                     /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(6443..6547)
                     /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 gattacaagg acgacgatga caaggacagc agcgctgccc ccacgaacgc
      961 cagcaattgc actgatgcct tggcgtactc aagttgctcc ccagcaccca gccccggttc
     1021 ctgggtcaac ttgtcccact tagatggcaa cctgtccgac ccatgcggtc cgaaccgcac
     1081 cgacctgggc gggagagaca gcctgtgccc tccgaccggc agtccctcca tgatcacggc
     1141 catcacgatc atggccctct actccatcgt gtgcgtggtg gggctcttcg gaaacttcct
     1201 ggtcatgtat gtgattgtca gatacaccaa gatgaagact gccaccaaca tctacatttt
     1261 caaccttgct ctggcagatg ccttagccac cagtaccctg cccttccaga gtgtgaatta
     1321 cctaatggga acatggccat ttggaaccat cctttgcaag atagtgatct ccatagatta
     1381 ctataacatg ttcaccagca tattcaccct ctgcaccatg agtgttgatc gatacattgc
     1441 agtctgccac cctgtcaagg ccttagattt ccgtactccc cgaaatgcca aaattatcaa
     1501 tgtctgcaac tggatcctct cttcagccat tggtcttcct gtaatgttca tggctacaac
     1561 aaaatacagg caaggttcca tagattgtac actaacattc tctcatccaa cctggtactg
     1621 ggaaaacctg ctgaagatct gtgttttcat cttcgccttc attatgccag tgctcatcat
     1681 taccgtgtgc tatggactga tgatcttgcg cctcaagagt gtccgcatgc tctctggctc
     1741 caaagaaaag gacaggaatc ttcgaaggat caccaggatg gtgctggtgg tggtggctgt
     1801 gttcatcgtc tgctggactc ccattcacat ttacgtcatc attaaagcct tggttacaat
     1861 cccagaaact acgttccaga ctgtttcttg gcacttctgc attgctctag gttacacaaa
     1921 cagctgcctc aacccagtcc tttatgcatt tctggatgaa aacttcaaac gatgcttcag
     1981 agagttctgt atcccaacct cttccaacat tgagcaacaa aactccactc gaattcgtca
     2041 gaacactaga gaccacccct ccacggccaa tacagtggat agaactaatc atcagctaga
     2101 aaatctggaa gcagaaactg ctccgttgcc ctagctcgag tctagagggc ccgtttaaac
     2161 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2221 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2281 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2341 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2401 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2521 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2641 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2761 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2821 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2881 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2941 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3061 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3121 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3181 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3241 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3301 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3361 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3421 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3481 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3541 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3601 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3661 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3721 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3781 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3841 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3901 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3961 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4021 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4081 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4141 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4201 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4261 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4321 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4381 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5341 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6601 tcccctatgg t
//
