LOCUS       V046328                 7914 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046328
VERSION     V046328
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..2594
                     /label="Entpd4(NM_026174)"
                     /note="Entpd4(NM_026174)"
                     /gene="Entpd4"
     CDS             2601..2666
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2690..3278
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3344..3392
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3594..4022
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4036..4365
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4432..5226
                     /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    5402..5523
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5572..5588)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5596..5612
                     /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(5620..5650)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5665..5686
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5974..6562)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6733..7593)
                     /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(7594..7698)
                     /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 ccaccatggg gaggattggc atttcctgtc
      781 tctttcctgc ctcttggcat tttagcatct ctccagtggg ctgtcctcga attctgaaca
      841 ccaatttacg acaaatcgtt gtcattagca tcctggctgc agctgtctcc cttttatact
      901 tctctgttgt cataatccgc agcaagtatg ggtggctgtc aaaggacaag aaatttcaaa
      961 ggtacttggc ccgagtcaca gacgttgagg ccacagacac caacaacccc agcgtgaact
     1021 atggcatcgt ggtggactgc ggcagcagtg ggtctcggat atttgtctac tgctggcctc
     1081 ggcacaatgg caaccctcac gatctgctgg acatcagaca gatgagggac aaaaaccgga
     1141 agccagtggt gatgaagatt aagcccggca tctcagagtt tgctacctct ccagaaaaag
     1201 tcagcgacta catttctccg cttctgagct ttgctgcaga acatgtgcct cgggcaaaac
     1261 acaaagagac acctctctac attctctgca cagctggaat gagagtcctt cctgaaagcc
     1321 agcagaaagc catcctagag gacctcctga ccgacatccc tgtgcactat gatttcctgt
     1381 tttctgactc ccatgccgaa gtcatctcag gaaaacaaga aggtgtgtat gcttggatcg
     1441 gcattaattt tgtcctcgga cggtttgagc atattgagga ggatgacgag gcggttgtgg
     1501 aagtcaacat tccgggcagc gagagcagcg aggccatcgt gcggaaaagg acagctggtg
     1561 tcctcgacat gggaggcgtg tctacccaga tagcgtacga agtcccccaa actgtaagct
     1621 ttgcctcctc gcagcaggaa gaagtagcta aaaacctgtt agctgaattc aacctggggt
     1681 gcgatgtcca ccagactgag catgtgtacc gcgtctacgt ggccacgttt cttgggtttg
     1741 gtggtaatgc tgcccggcag agatatgaag accgactatt tgccagcaca gttcagaaaa
     1801 acaggctcct gggtaaacag actggtctga ctcctgatgc tccactactg gatccctgct
     1861 tgcctctgga cattaaagat gagatccagc aaaacgggca gaccctgtac cttcagggga
     1921 caggagactt tgacctgtgt cgagagaccc tgcagccttt catgaacaaa accaatgaga
     1981 cccagacttc cctcaatggc gtctaccagc ctccaatcca cttccagaac agtgaattct
     2041 acggcttctc tgagttctac tactgcaccg aggatgtctt gcgaatgggg ggagactaca
     2101 atgctgctag attcactcaa gctgccaagg attactgtgc aacaaagtgg tcgatcctgc
     2161 gggaacgctt tgaccgagga ctctatgcct ctcatgccga cctccatcga ctgaagtatc
     2221 agtgtttcaa atcagcctgg atgttcgagg tgttccacaa aggcttctcc tttcctgtca
     2281 cctacaaaaa cctgaagacg gccttgcagg tgtatgacaa ggaagtacag tggaccctgg
     2341 gggccatcct ttaccggacc cgcttcctgc ccttgagaga catccggcag gaggtgttcc
     2401 gagctggcca cgcgcactgg cggggcgtgt ccttcgtcta caaccactat ctgttctccg
     2461 gttgcttcct ggtcgtcctt ctgtccatcc ttctctacct gctgcggctg cggcgcatcc
     2521 accgcagggc gccccgcact ggctctctgt ggatggagga aggcctgccc tcccagaagg
     2581 gccctgggcc cttgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     2641 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3061 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3241 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3301 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3361 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3421 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3481 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3541 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3661 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3721 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3781 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3901 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3961 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4021 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4381 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4441 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4501 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4561 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4621 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4681 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4741 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4801 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4861 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4921 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4981 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5041 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5101 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5161 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5221 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5281 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5341 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5401 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5521 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5581 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5641 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5701 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5761 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7021 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7741 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7801 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7861 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
