LOCUS       V036543                 7821 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036543
VERSION     V036543
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..2501
                     /label="ZNF326(NM_182976)"
                     /note="ZNF326(NM_182976)"
                     /gene="ZNF326"
     CDS             2508..2573
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2597..3185
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3251..3299
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3501..3929
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3943..4272
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4339..5133
                     /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    5309..5430
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5479..5495)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5503..5519
                     /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(5527..5557)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5572..5593
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5881..6469)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6640..7500)
                     /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(7501..7605)
                     /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 ccaccatgga cttcgaggac gattacacac
      781 actccgcctg caggaatact tatcagggct ttaatggaat ggatcgtgat tatggccctg
      841 gatcttatgg agggatggat cgtgactatg gccatggatc ctatgggggt cagagatcca
      901 tggattccta cctaaaccag tcatatggca tggacaatca cagtggtggt ggtgggggta
      961 gcaggtttgg accttatgag tcttacgact ccaggtcttc tctgggtggg cgagatctgt
     1021 acagatctgg ctatggtttt aatgaacccg aacaaagccg cttcggaggt agttatggtg
     1081 gtcgatttga gagctcctac cggaatagcc ttgactcttt cggaggtaga aaccagggcg
     1141 ggtctagctg ggaagcacct tactcccgtt caaaattgag gcctgggttt atggaggaca
     1201 gaggaagaga gaattactct tcctacagca gtttttcttc accccatatg aagcctgcac
     1261 ctgtaggctc tcgggggaga ggaacgcctg cttatcctga aagtacgttt ggaagcagaa
     1321 actatgatgc ttttggagga ccatcaacag gcagaggccg aggccgagga catatgggtg
     1381 attttggaag cattcataga cccggaattg ttgttgacta tcaaaacaaa tccaccaatg
     1441 tgacagttgc tgctgcaaga ggaataaaga gaaaaatgat gcagccattt aataagccca
     1501 gtggaacctt tatcaagaaa cccaaactag caaaacctat ggagaagata agcctcagca
     1561 aatcacccac aaaaactgat cctaaaaatg aagaggaaga aaagcggcga attgaggctc
     1621 ggcgagagaa acaaaggcgc agaagagaaa aaaacagtga gaaatacgga gatggataca
     1681 gaatggcatt tacatgttca ttttgtaaat ttcgaacatt tgaagaaaaa gatattgaac
     1741 tgcatctgga aagttcttca catcaggaaa cattagatca tatacagaaa caaactaaat
     1801 ttgataaagt agttatggag tttttgcatg agtgtatggt gaataaattc aagaaaacat
     1861 ctattcgtaa gcaacagaca aataatcaaa cagaagtagt taaaataatt gaaaaagatg
     1921 ttatggaagg tgttactgta gatgatcaca tgatgaaggt agagacagtt cattgcagcg
     1981 cttgcagtgt ttatatccct gctttacata gttcagttca gcagcactta aaatctcctg
     2041 atcatatcaa agggaagcag gcttataagg aacaaataaa aagagagagt gtcttgactg
     2101 ctacaagcat tttaaataat ccaatagtga aggcgcgata tgaacgtttt gttaagggtg
     2161 agaatccttt tgaaattcaa gaccattctc aggatcagca aatagaagga gatgaggagg
     2221 atgaagagaa gattgatgaa cctattgaag aagaggagga tgaagatgag gaagaagaag
     2281 cagaggaagt gggggaagta gaggaagtgg aagaagtaga ggaagtgaga gaaggaggaa
     2341 tagagggcga gggaaatata cagggagtag gggaaggagg ggaagtaggg gtagtgggag
     2401 aagtagaggg agtgggggaa gtagaggaag tagaggaatt agaggaagag acagcaaagg
     2461 aagaacctgc tgacttccct gttgagcaac ctgaagaaaa tggatccgac tacaaagacc
     2521 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     2581 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3001 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3181 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3241 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     3301 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3361 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3421 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     3481 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3541 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3601 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3661 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3721 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3781 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3841 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3901 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4081 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4141 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4201 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4261 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4321 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4381 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4441 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4501 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4561 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4621 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4681 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4741 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4801 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4861 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4921 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4981 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5041 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5101 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     5161 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5221 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5281 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5341 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5401 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5461 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5521 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5581 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6061 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6301 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6481 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6541 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7621 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7681 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7741 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7801 tgcatgaaga atctgcttag g
//
