LOCUS       V045365                 7534 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045365
VERSION     V045365
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             706..2190
                     /label="NUFIP1(NM_012345)"
                     /note="NUFIP1(NM_012345)"
                     /gene="NUFIP1"
     CDS             2197..2286
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2310..2898
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2964..3012
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3214..3642
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3656..3985
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4052..4846
                     /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    5022..5143
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5192..5208)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5216..5232
                     /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(5240..5270)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5285..5306
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5594..6182)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6353..7213)
                     /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(7214..7318)
                     /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 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc tgagccgact
      721 agtgatttcg agactcctat cgggtggcat gcgtctcccg agctgactcc cacgttaggg
      781 cccctgagcg acactgcccc gccgcgggac agctggatgt tctgggcaat gctgccgcca
      841 ccgccaccac cacttacgtc ctcgcttccc gcagccgggt caaagccttc ctctgagtcg
      901 cagcccccca tggaggccca gtctctcccc ggggctccgc cccccttcga cgcccagatt
      961 cttcccgggg cgcaaccccc cttcgacgcc cagtctcccc ttgattctca gcctcaaccc
     1021 agcggccagc cttggaattt ccatgcttcc acatcgtggt attggagaca gtcttctgat
     1081 aggtttcctc ggcatcagaa gtccttcaac cctgcagtta aaaattctta ttatccacga
     1141 aagtatgatg caaaattcac agacttcagc ttacctccca gtagaaaaca gaaaaaaaag
     1201 aaaagaaagg aaccagtttt tcactttttt tgtgatacct gtgatcgtgg ttttaaaaat
     1261 caagaaaagt atgacaaaca catgtctgaa catacaaaat gccctgaatt agattgctct
     1321 tttactgcac acgagaagat tgtccagttc cattggagaa atatgcatgc tcctggcatg
     1381 aagaagatca agttagacac tccagaggaa attgcacggt ggagggaaga aagaaggaaa
     1441 aactatccaa ctctggccaa tattgaaagg aagaagaagt taaaacttga aaaggagaag
     1501 agaggagcag tattgacaac aacacaatat ggcaagatga aggggatgtc cagacattca
     1561 caaatggcaa agatcagaag tcctggcaag aatcacaaat ggaaaaacga caattctaga
     1621 cagagagcag tcactggatc aggcagtcac ttgtgtgatt tgaagctaga aggtccaccg
     1681 gaggcaaatg cagatcctct tggtgttttg ataaacagtg attctgagtc tgataaggag
     1741 gagaaaccac aacattctgt gatacccaag gaagtgacac cagccctatg ctcactaatg
     1801 agtagctatg gcagtctttc agggtcagag agtgagccag aagaaactcc catcaagact
     1861 gaagcagacg ttttggcaga aaaccaggtt cttgatagca gtgctcctaa gagtccaagt
     1921 caagatgtta aagcaactgt tagaaatttt tcagaagcca agagtgagaa ccgaaagaaa
     1981 agctttgaaa aaacaaaccc taagaggaaa aaagattatc acaactatca aacgttattc
     2041 gaaccaagaa cacaccatcc atatctcttg gaaatgcttc tagctccgga cattcgacat
     2101 gaaagaaatg tgattttgca gtgtgttcgg tacatcatca aaaaagactt ttttggactg
     2161 gatactaatt ctgcgaaaag taaagatgta ctcgaggaac aaaaactaat atcagaggaa
     2221 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     2281 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     2341 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2401 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2461 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2521 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2581 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2641 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2701 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2761 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2821 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2881 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     2941 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     3001 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3061 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3121 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3181 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3361 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3421 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3481 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3541 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     3601 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3661 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3841 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3901 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3961 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4021 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4081 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4141 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4201 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4261 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4321 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4381 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4441 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4501 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4561 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4621 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4681 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4741 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4801 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4861 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4921 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4981 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5161 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5761 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6001 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7321 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7381 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7441 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7501 cttgaccgac aattgcatga agaatctgct tagg
//
