LOCUS       V027549                 7902 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027549
VERSION     V027549
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..2582
                     /label="ZNF527(NM_032453)"
                     /note="ZNF527(NM_032453)"
                     /gene="ZNF527"
     CDS             2589..2654
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2678..3266
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3332..3380
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3582..4010
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4024..4353
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4420..5214
                     /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    5390..5511
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5560..5576)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5584..5600
                     /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(5608..5638)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5653..5674
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5962..6550)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6721..7581)
                     /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(7582..7686)
                     /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 ccaccatggc tgtggggctt tgtaaagcca
      781 tgtcccaggg gttggtgacc ttcagagatg tggcgctaga cttttcccaa gaagagtggg
      841 aatggctgaa gccatctcag aaggatttat acagagatgt catgttggag aactacagga
      901 acttggtatg gcttggactc tccatttcta agcccaacat gatctcctta ctggagcaag
      961 ggaaggaacc gtggatggtg gagagaaaga tgtcacaggg tcactgtgca gactgggagt
     1021 cttggtgtga aattgaggaa ttatctccaa aatggttcat tgatgaagat gaaatatccc
     1081 aggagatggt aatggaaagg ctagcaagtc atggccttga atgctccagt ttcagagaag
     1141 cctggaaata taagggtgaa tttgagctac atcagggaaa tgcggagagg catttcatgc
     1201 aagtgacagc tgttaaggaa atctctactg ggaaaagaga caatgaattt agtaattctg
     1261 ggagaagcat acccctgaaa tcagtatttt taacacaaca gaaagttcct accatacagc
     1321 aagtacataa atttgatatt tatgataaac tcttccccca aaattcagtc ataattgaat
     1381 ataaaagact ccatgctgag aaggaatctt tgataggtaa tgaatgtgaa gaattcaacc
     1441 agagtacgta ccttagtaaa gatataggaa ttcctcctgg ggagaaacct tatgaaagtc
     1501 atgatttttc aaagctctta agtttccact cattatttac tcaacatcag accactcatt
     1561 ttggaaaatt accccatgga tacgatgaat gtggtgatgc ctttagctgt tactcattct
     1621 ttactcaacc tcagagaatt cacagtggag aaaaaccata tgcatgcaat gactgtggaa
     1681 aagcctttag ccacgacttc tttctcagtg aacatcaaag aactcatatt ggggagaaac
     1741 cttatgaatg taaggaatgt aacaaagctt tcagacagag tgctcacctt gctcaacatc
     1801 agaggatcca cactggagag aaaccgtttg cgtgcaatga atgtgggaag gcctttagcc
     1861 gttatgcctt ccttgttgaa catcagagaa ttcacacagg tgagaaacca tatgaatgta
     1921 aagaatgtaa taaagccttc agacagagtg ctcaccttaa tcaacatcag aggattcaca
     1981 ctggagagaa accctatgaa tgtaatcagt gtggaaaagc cttcagcaga cgcatagccc
     2041 ttactctaca tcaaagaatt cacacaggag agaaaccctt caaatgtagt gaatgtggga
     2101 agacctttgg ctatcgctca cacctgaatc aacatcagag aattcatacc ggagaaaagc
     2161 cctatgaatg catcaaatgt gggaagtttt ttaggactga ctcacaactt aatcgacatc
     2221 atagaattca cactggagag agaccatttg aatgcagtaa atgtgggaaa gccttcagtg
     2281 atgctttagt tctaattcac cataagagaa gtcatgcagg agagaaaccc tatgaatgta
     2341 acaaatgtgg aaaggccttc agttgtggct catatcttaa tcaacatcaa agaattcata
     2401 ctggagagaa accctatgaa tgtagtgaat gtgggaaggc ttttcatcag atcttgtccc
     2461 taagactaca ccagagaatt cacgctggag aaaaacctta taaatgtaac gaatgtggga
     2521 ataattttag ctgtgtctca gcccttagac gacatcagag aattcataat agagaaacgc
     2581 tcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2641 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     2701 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2761 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2821 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2881 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2941 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3001 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3061 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3121 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3181 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3241 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     3301 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3361 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3421 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3481 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3541 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3601 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3661 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3721 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3781 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3841 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3901 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3961 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4021 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4141 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4201 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4261 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4321 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4381 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4441 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4501 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4561 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4621 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4681 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4741 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4801 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4861 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4921 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4981 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5041 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5101 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5161 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5221 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5281 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5341 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5521 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5581 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5641 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5701 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5761 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6121 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6361 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7681 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7741 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7801 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7861 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
