LOCUS       V045261                 7971 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045261
VERSION     V045261
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..2651
                     /label="Acoxl(NM_028765)"
                     /note="Acoxl(NM_028765)"
                     /gene="Acoxl"
     CDS             2658..2723
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2747..3335
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3401..3449
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3651..4079
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4093..4422
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4489..5283
                     /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    5459..5580
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5629..5645)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5653..5669
                     /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(5677..5707)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5722..5743
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6031..6619)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6790..7650)
                     /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(7651..7755)
                     /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 ccaccatgac aggatggagt ggactgttta
      781 tagaagtccc tctgtataga ttggcatcca aagtgtgctg tcatctgcgg ggaccacagg
      841 gatttcttca gagagcctgc cagggagtgt gttgtctgcc aagggactcc agagcagctg
      901 gaatcatgga agaaaaaagg aagagcttca tcagccaaat ccttatcctg ggagaagtgc
      961 tctgcatggt ggatgtgtcc atgtctataa agtgtggaat cctttttttg ttatttggtg
     1021 gtgcaatcag caatcttgga agtcctgaac atgttaccaa gtggttttgg ccactcaagg
     1081 agcagaagta tactgggatg tttgcaatga ctgagagggg ccatgggagc aacgtgagag
     1141 ggatccagac tgaggccacc tttgacctcg acaatcagga gtttgtgatt gacatgccgt
     1201 gtgagaacgc ccacaagatg tacatcggca atgccatgca tgggaactac gctgctgtct
     1261 tcgcccaact catcattgag ggaaagtctc aagggcccca ctgttttatt gtccctatca
     1321 gggatgaaaa tggaaacttg tacccaggag tgacagctat tgacatgatg cacaaagagg
     1381 gtatgaatgg tgtggataac ggaattttga tatttgacaa ggttcggatt cccagagaga
     1441 acctgctgga taagtttggt tctgtgactc cagatggaca ataccattca cctattcaaa
     1501 gcaagaacgc aagattcaat gcgatcctgg caacactgac tccttcaaga ctagcagtga
     1561 ctttccaagc tttgggcgcc atgaagcttg gactgatgat agccatccgc tacagccaca
     1621 gccgaaggca atttgggccc aaggacaagg aagaagtgaa gatcattgag caccaaatgc
     1681 aggcactcag gctgatgtct cacctggcca cagccctggc cgtgaccttc acaagcaggc
     1741 atgcagacga catcctggat gaagacatct tccagggaag ggcgctcacc aacagtcgct
     1801 cgctacaggc actgatggct ggactgaaag cctacagtac atgggagact gttagctgcc
     1861 tacaggactg tcgtgaatgc accggaggca tgggttacat gatggaaacc agaatctcag
     1921 acctgaaatg tgacacagat gtgtttgtaa cctttgaagg tgacaatgtg gtcatgcttc
     1981 aggttgtggc ccgggaactg ctggcccagt acagcaagca gcataagaaa aacctgctct
     2041 tgggtgtgat ccagaactgg acagcaactg ccggggacaa gctgagaacc agtttcctgg
     2101 cgtttaacac tgacacggta ggatgtcttg cctttctgct gaaagcagtg aattttcgag
     2161 aaagggttct tcagcggagt ttggtgtcca gaatttacta taaggtggtg accaagaagg
     2221 gggacttctt cagtgcctgg aactcctgta tgcaccatgt cacctcattg tctctggcac
     2281 acattcacag agttgcacta gagcagttca cgacggcagt gaggcaatgt cccaacaggg
     2341 aggaccaggc tttgctaatg aagttttgtc tgctgtatgg aacgaagctg gtgttccagg
     2401 agcgaggctg gtacttagaa cataaatacc tgacccccaa ggccagcatg ctgatcaggg
     2461 ctcagttgct gaatttgtgc gagtccgtga aggatgacgc cctgaaggtg atctctgcct
     2521 tcaacattcc acacatcacc atccgtgcac caaaaacagg aatccccaac cccggggctg
     2581 ccgaggccgc ctacccggca cccatgcaac cgctcgtgcg ggatgcggcg cgcgctcaac
     2641 tggccaagct gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2701 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2761 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2821 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2881 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2941 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3001 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3061 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3121 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3181 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3241 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3301 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3361 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3421 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3481 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3541 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3601 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3661 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3721 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3781 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3841 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3901 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3961 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4021 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4081 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4141 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4261 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4321 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4381 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4441 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4501 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4561 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4621 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4681 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4741 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4801 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4861 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4921 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4981 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5041 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5101 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5161 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5221 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5281 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5341 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5401 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5461 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5521 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5581 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5641 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5701 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5821 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7801 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7861 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7921 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
