LOCUS       V025142                 9348 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025142
VERSION     V025142
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4028
                     /label="ADCY2(NM_020546)"
                     /note="ADCY2(NM_020546)"
                     /gene="ADCY2"
     CDS             4035..4100
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4124..4712
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4778..4826
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5028..5456
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5470..5799
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5866..6660
                     /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    6836..6957
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7006..7022)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7030..7046
                     /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(7054..7084)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7099..7120
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7408..7996)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8167..9027)
                     /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(9028..9132)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtg gcaggaggcg atgcggcgcc
      781 gccgctacct gcgggaccgc tccgaggagg cggcgggcgg cggagacggg ctgccgcggt
      841 cccgggactg gctctacgag tcctactact gcatgagcca gcagcacccg ctcatcgtct
      901 tcctgctgct catcgtcatg ggctcctgcc tcgccctgct cgccgtcttc ttcgcgctcg
      961 ggctggaagt tgaagaccat gtggcgtttc taataacagt tccaactgcc ctggcgattt
     1021 tctttgcgat atttatcctg gtctgcatcg agtctgtgtt taagaagctg ctgcgcctct
     1081 tctcgttggt gatatggata tgccttgttg ccatgggata cctgttcatg tgttttggag
     1141 gcaccgtctc tccctgggac caggtatcgt tcttcctctt catcatcttc gtggtgtaca
     1201 ccatgctgcc cttcaacatg cgagacgcca tcattgccag cgtcctcacc tcctcctccc
     1261 acaccatcgt gcttagcgtc tgcctgtctg caacaccggg aggcaaggag cacctggtct
     1321 ggcagatcct ggccaatgtg atcattttca tctgtgggaa cctggcggga gcctaccata
     1381 agcacctcat ggaactcgct cttcagcaaa catatcagga cacctgtaat tgcatcaagt
     1441 cgcggatcaa gttggaattt gaaaaacgtc aacaggagcg gcttctgctc tccctgctgc
     1501 cggcccacat cgccatggag atgaaagcgg agatcatcca gaggctgcag ggccccaagg
     1561 cgggccagat ggagaacaca aataacttcc acaacctgta tgtgaagcgg catacaaacg
     1621 tgagcatctt atacgctgac atcgttggct ttacccggct ggcaagtgac tgctccccgg
     1681 gagaactagt ccacatgctg aatgagctct ttggaaagtt tgatcaaatt gcaaaggaga
     1741 atgaatgcat gagaattaaa attttaggag actgctacta ctgtgtatct ggactcccta
     1801 tatctctccc taaccatgcc aagaactgtg tgaaaatggg gctggacatg tgtgaagcca
     1861 taaagaaagt gagggatgct actggagttg atatcaacat gcgcgtgggc gtgcattctg
     1921 ggaatgtcct gtgtggcgtg attggtctgc agaagtggca atatgatgtg tggtcacatg
     1981 atgtgacctt ggccaaccac atggaagctg gaggggtccc tggacgtgtt cacatttctt
     2041 ctgtcaccct ggagcacttg aatggcgctt ataaagtgga ggagggagat ggtgacatta
     2101 gggacccata tttaaaacag cacctggtga aaacctactt tgtgatcaac cccaagggag
     2161 aacgacggag cccccagcat ctcttcagac ctcgccacac ccttgatgga gccaaaatga
     2221 gggcctcggt ccgcatgacc cggtacttgg agtcctgggg ggcagccaag ccctttgcac
     2281 acctacatca cagggacagc atgaccacag agaacggcaa gatcagcacc acggatgtac
     2341 ccatgggtca gcataatttt caaaatcgca ccttaagaac caagtcacaa aagaagagat
     2401 ttgaagaaga attgaatgaa aggatgattc aagcaattga tgggattaat gcacagaagc
     2461 aatggctcaa gtctgaagac attcagagaa tctcactgct tttctataac aaagtactag
     2521 aaaaagagta ccgggccacg gcactgccag cgttcaagta ttatgtgact tgtgcctgtc
     2581 tcatattctt ctgcatcttc attgtgcaga ttctcgtgct gccaaaaacg tccgtcctgg
     2641 gcatctcctt tggggctgcg tttctcttgc tggccttcat cctcttcgtc tgctttgctg
     2701 gacagcttct gcaatgcagc aaaaaagcct ctcccctgct catgtggctt ttgaagtcct
     2761 cgggcatcat tgccaaccgc ccctggccac ggatctctct cacgatcatc accacagcca
     2821 tcatattaat gatggccgtg ttcaacatgt ttttcctgag tgactcagag gaaacaatcc
     2881 ctccaactgc caacacaaca aacacaagct tttcagcctc aaataatcag gtggcgattc
     2941 tgcgtgcgca gaatttattt ttcctcccgt actttatcta cagctgcatt ctgggactga
     3001 tatcctgttc cgtgttcctg cgggtaaact atgagctgaa gatgttgatc atgatggtgg
     3061 ccttggtggg ctacaacacc atcctactcc acacccacgc ccacgtcctg ggcgactaca
     3121 gccaggtctt atttgagaga ccaggcattt ggaaagacct gaagaccatg ggctctgtgt
     3181 ctctctctat attcttcatc acactgcttg ttctgggtag acagaatgaa tattactgta
     3241 ggttagactt cttatggaag aacaaattca aaaaagagcg ggaggagata gagaccatgg
     3301 agaacctgaa ccgcgtgctg ctggagaacg tgcttcccgc gcacgtggct gagcacttcc
     3361 tggccaggag cctgaagaat gaggagctat accaccagtc ctatgactgc gtctgcgtca
     3421 tgtttgcctc cattccggat ttcaaagaat tttatacaga atccgacgtg aacaaggagg
     3481 gcttggaatg ccttcggctc ctgaacgaga tcatcgctga ctttgatgat cttctttcca
     3541 agccaaaatt cagtggagtt gaaaagatta agaccattgg cagcacatac atggcagcaa
     3601 caggtctgag cgctgtgccc agccaggagc actcccagga gcccgagcgg cagtacatgc
     3661 acattggcac catggtggag tttgcttttg ccctggtagg gaagctggat gccatcaaca
     3721 agcactcctt caacgacttc aaattgcgag tgggtattaa ccatggacct gtgatagctg
     3781 gtgtgattgg agctcagaag ccacaatatg atatctgggg caacactgtc aatgtggcca
     3841 gtaggatgga cagcaccgga gtcctggaca aaatacaggt taccgaggag acgagcctcg
     3901 tcctgcagac cctcggatac acgtgcacct gtcgaggaat aatcaacgtg aaaggaaagg
     3961 gggacctgaa gacgtacttt gtaaacacag aaatgtcaag gtccctttcc cagagcaacg
     4021 tggcatccgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     4081 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     4141 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4201 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4261 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4321 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4381 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4441 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4501 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4561 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4621 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4681 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     4741 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4801 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4861 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4921 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4981 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     5041 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5101 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5161 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     5221 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5281 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5341 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     5401 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     5461 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5521 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5641 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5701 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5761 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5821 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5881 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5941 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6001 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6061 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6121 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6181 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6241 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6301 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6361 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6421 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6481 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6541 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6601 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6661 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6721 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6781 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6841 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6901 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6961 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7021 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7081 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7141 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7201 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7261 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7321 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7381 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7441 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7501 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7561 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7621 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7681 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7741 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7801 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7861 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7921 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7981 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8041 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8101 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     9181 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     9241 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     9301 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
