LOCUS       V018912                 9464 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018912
VERSION     V018912
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"
     CDS             671..4144
                     /label="NRIP1(NM_003489)"
                     /note="NRIP1(NM_003489)"
                     /gene="NRIP1"
     CDS             4151..4216
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4240..4828
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4894..4942
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5144..5572
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5586..5915
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5982..6776
                     /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    6952..7073
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7122..7138)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7146..7162
                     /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(7170..7200)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7215..7236
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7524..8112)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8283..9143)
                     /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(9144..9248)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atgactcatg gagaagagct tggctctgat gtgcaccagg attctattgt
      721 tttaacttac ctagaaggat tactaatgca tcaggcagca gggggatcag gtactgccgt
      781 tgacaaaaag tctgctgggc ataatgaaga ggatcagaac tttaacattt ctggcagtgc
      841 atttcccacc tgtcaaagta atggtccagt tctcaataca catacatatc aggggtctgg
      901 catgctgcac ctcaaaaaag ccagactgtt gcagtcttct gaggactgga atgcagcaaa
      961 gcggaagagg ctgtctgatt ctatcatgaa tttaaacgta aagaaggaag ctttgctagc
     1021 tggcatggtt gacagtgtgc ctaaaggcaa acaggatagc acattactgg cctctttgct
     1081 tcagtcattc agctctaggc tgcagactgt tgctctgtca caacaaatca ggcagagcct
     1141 caaggagcaa ggatatgccc tcagtcatga ttctttaaaa gtggagaagg atttaaggtg
     1201 ctatggtgtt gcatcaagtc acttaaaaac tttgttgaag aaaagtaaag ttaaagatca
     1261 aaagcctgat acgaatcttc ctgatgtgac taaaaacctc atcagagata ggtttgcaga
     1321 gtctcctcat catgttggac aaagtggaac aaaggtcatg agtgaaccgt tgtcatgtgc
     1381 tgcaagatta caggctgttg caagcatggt ggaaaaaagg gctagtcctg ccacctcacc
     1441 taaacctagt gttgcttgta gccagttagc attacttctg tcaagcgaag cccatttgca
     1501 gcagtattct cgagaacacg ctttaaaaac gcaaaatgca aatcaagcag caagtgaaag
     1561 acttgctgct atggccagat tgcaagaaaa tggccagaag gatgttggca gttaccagct
     1621 cccaaaagga atgtcaagcc atcttaatgg tcaggcaaga acatcatcaa gcaaactgat
     1681 ggctagcaaa agtagtgcta cagtgtttca aaatccaatg ggtatcattc cttcttcccc
     1741 taaaaatgca ggttataaga actcactgga aagaaacaat ataaaacaag ctgctaacaa
     1801 tagtttgctt ttacatcttc ttaaaagcca gactatacct aagccaatga atggacacag
     1861 tcacagtgag agaggaagca tttttgagga aagtagtaca cctacaacta ttgatgaata
     1921 ttcagataac aatcctagtt ttacagatga cagcagtggt gatgaaagtt cttattccaa
     1981 ctgtgttccc atagacttgt cttgcaaaca ccgaactgaa aaatcagaat ctgaccaacc
     2041 tgtttccctg gataacttca ctcaatcctt gctaaacact tgggatccaa aagtcccaga
     2101 tgtagatatc aaagaagatc aagatacctc aaagaattct aagctaaact cacaccagaa
     2161 agtaacactt cttcaattgc tacttggcca taagaatgaa gaaaatgtag aaaaaaacac
     2221 cagccctcag ggagtacaca atgatgtgag caagttcaat acacaaaatt atgcaaggac
     2281 ttctgtgata gaaagcccca gtacaaatcg gactactcca gtgagcactc cacctttact
     2341 tacatcaagc aaagcagggt ctcccatcaa tctctctcaa cactctctgg tcatcaaatg
     2401 gaattcccca ccatatgtct gcagtactca gtctgaaaag ctaacaaata ctgcatctaa
     2461 ccactcaatg gaccttacaa aaagcaaaga cccaccagga gagaaaccag cccaaaatga
     2521 aggtgcacag aactctgcaa cgtttagtgc cagtaagctg ttacaaaatt tagcacaatg
     2581 tggaatgcag tcatccatgt cagtggaaga gcagagaccc agcaaacagc tgttaactgg
     2641 aaacacagat aaaccgatag gtatgattga tagattaaat agccctttgc tctcaaataa
     2701 aacaaatgca gttgaagaaa ataaagcatt tagtagtcaa ccaacaggtc ctgaaccagg
     2761 gctttctggt tctgaaatag aaaatctgct tgaaagacgt actgtcctcc agttgctcct
     2821 ggggaacccc aacaaaggga agagtgaaaa aaaagagaaa actcccttaa gagatgaaag
     2881 tactcaggaa cactcagaga gagctttaag tgaacaaata ctgatggtga aaataaaatc
     2941 tgagccttgt gatgacttac aaattcctaa cacaaatgtg cacttgagcc atgatgctaa
     3001 gagtgcccca ttcttgggta tggctcctgc tgtgcagaga agcgcacctg ccttaccagt
     3061 gtccgaagac tttaaatcgg agcctgtttc acctcaggat ttttctttct ccaagaatgg
     3121 tctgctaagt cgattgctaa gacaaaatca agatagttac ctggcagatg attcagacag
     3181 gagtcacaga aataatgaaa tggcacttct agaatcaaag aatctttgca tggtccctaa
     3241 gaaaaggaag ctttatactg agccattaga aaatccattt aaaaagatga aaaacaacat
     3301 tgttgatgct gcaaacaatc acagtgcccc agaagtactg tatgggtcct tgcttaacca
     3361 ggaagagctg aaatttagca gaaatgatct tgaatttaaa tatcctgctg gtcatggctc
     3421 agccagcgaa agtgaacaca ggagttgggc cagagagagc aaaagcttta atgttctgaa
     3481 acagctgctt ctctcagaaa actgtgtgcg agatttgtcc ccgcacagaa gtaactctgt
     3541 ggctgacagt aaaaagaaag gacacaaaaa taatgtgacc aacagcaaac ctgaatttag
     3601 catttcttct ttaaatggac tgatgtacag ttccactcag cccagcagtt gcatggataa
     3661 caggacattt tcatacccag gtgtagtaaa aactcctgtg agtcctactt tccctgagca
     3721 cttgggctgt gcagggtcta gaccagaatc tgggcttttg aatgggtgtt ccatgcccag
     3781 tgagaaagga cccattaagt gggttatcac tgatgcggag aagaatgagt atgaaaaaga
     3841 ctctccaaga ttgaccaaaa ccaacccaat actatattac atgcttcaaa aaggaggcaa
     3901 ttctgttacc agtcgagaaa cacaagacaa ggacatttgg agggaggctt catctgctga
     3961 aagtgtctca caggtcacag ccaaagaaga gttacttcct actgcagaaa cgaaagcttc
     4021 tttctttaat ttaagaagcc cttacaatag ccatatggga aataatgctt ctcgcccaca
     4081 cagcgcaaat ggagaagttt atggacttct gggaagcgtg ctaacgataa agaaagaatc
     4141 agaactcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4201 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     4261 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4321 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4381 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4441 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4501 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4561 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4621 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4681 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4741 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4801 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4861 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4921 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4981 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5041 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5101 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5281 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     5341 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5461 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     5521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5581 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5641 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5761 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5821 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5881 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5941 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6001 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6061 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6121 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     6181 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6241 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6301 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6361 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6421 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6481 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6541 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6601 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6661 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6721 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6781 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6841 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6901 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6961 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7021 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7081 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7141 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7681 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7921 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8161 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8221 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8281 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8341 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8401 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8461 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8521 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8581 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8641 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8701 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8761 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8821 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8881 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8941 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9001 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9061 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9121 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9181 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9241 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     9301 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9361 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9421 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
