LOCUS       V023312                 7803 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023312
VERSION     V023312
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..2477
                     /label="NT5E(NM_002526)"
                     /note="NT5E(NM_002526)"
                     /gene="NT5E"
     CDS             2490..2555
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2579..3167
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3233..3281
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3483..3911
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3925..4254
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4321..5115
                     /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    5291..5412
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5461..5477)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5485..5501
                     /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(5509..5539)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5554..5575
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5863..6451)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6622..7482)
                     /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(7483..7587)
                     /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 ctcggatccg ccaccatgtg tccccgagcc gcgcgggcgc
      781 ccgcgacgct actcctcgcc ctgggcgcgg tgctgtggcc tgcggctggc gcctgggagc
      841 ttacgatttt gcacaccaac gacgtgcaca gccggctgga gcagaccagc gaggactcca
      901 gcaagtgcgt caacgccagc cgctgcatgg gtggcgtggc tcggctcttc accaaggttc
      961 agcagatccg ccgcgccgaa cccaacgtgc tgctgctgga cgccggcgac cagtaccagg
     1021 gcactatctg gttcaccgtg tacaagggcg ccgaggtggc gcacttcatg aacgccctgc
     1081 gctacgatgc catggcactg ggaaatcatg aatttgataa tggtgtggaa ggactgatcg
     1141 agccactcct caaagaggcc aaatttccaa ttctgagtgc aaacattaaa gcaaaggggc
     1201 cactagcatc tcaaatatca ggactttatt tgccatataa agttcttcct gttggtgatg
     1261 aagttgtggg aatcgttgga tacacttcca aagaaacccc ttttctctca aatccaggga
     1321 caaatttagt gtttgaagat gaaatcactg cattacaacc tgaagtagat aagttaaaaa
     1381 ctctaaatgt gaacaaaatt attgcactgg gacattcggg ttttgaaatg gataaactca
     1441 tcgctcagaa agtgaggggt gtggacgtcg tggtgggagg acactccaac acatttcttt
     1501 acacaggcaa tccaccttcc aaagaggtgc ctgctgggaa gtacccattc atagtcactt
     1561 ctgatgatgg gcggaaggtt cctgtagtcc aggcctatgc ttttggcaaa tacctaggct
     1621 atctgaagat cgagtttgat gaaagaggaa acgtcatctc ttcccatgga aatcccattc
     1681 ttctaaacag cagcattcct gaagatccaa gcataaaagc agacattaac aaatggagga
     1741 taaaattgga taattattct acccaggaat tagggaaaac aattgtctat ctggatggct
     1801 cctctcaatc atgccgcttt agagaatgca acatgggcaa cctgatttgt gatgcaatga
     1861 ttaacaacaa cctgagacac acggatgaaa tgttctggaa ccacgtatcc atgtgcattt
     1921 taaatggagg tggtatccgg tcgcccattg atgaacgcaa caatggcaca attacctggg
     1981 agaacctggc tgctgtattg ccctttggag gcacatttga cctagtccag ttaaaaggtt
     2041 ccaccctgaa gaaggccttt gagcatagcg tgcaccgcta cggccagtcc actggagagt
     2101 tcctgcaggt gggcggaatc catgtggtgt atgatctttc ccgaaaacct ggagacagag
     2161 tagtcaaatt agatgttctt tgcaccaagt gtcgagtgcc cagttatgac cctctcaaaa
     2221 tggacgaggt atataaggtg atcctcccaa acttcctggc caatggtgga gatgggttcc
     2281 agatgataaa agatgaatta ttaagacatg actctggtga ccaagatatc aacgtggttt
     2341 ctacatatat ctccaaaatg aaagtaattt atccagcagt tgaaggtcgg atcaagtttt
     2401 ccacaggaag tcactgccat ggaagctttt ctttaatatt tctttcactt tgggcagtga
     2461 tctttgtttt ataccaactc gagtctagag actacaaaga ccatgacggt gattataaag
     2521 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     2581 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2641 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2701 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2761 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2821 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2881 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2941 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3001 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3061 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3121 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3181 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3241 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3301 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3361 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3421 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3481 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3841 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3901 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4261 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4321 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4381 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4441 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4501 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4561 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4621 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4681 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4741 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4801 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4861 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4921 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4981 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5041 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5101 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5161 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5221 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5281 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5341 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5401 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5461 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5521 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5581 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6301 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6421 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6481 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6541 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6601 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6661 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6721 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6781 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6841 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6901 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6961 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7021 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7081 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7141 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7201 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7261 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7321 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7381 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7441 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7501 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7561 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7621 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     7681 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7741 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7801 agg
//
