LOCUS       V025841                 8817 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025841
VERSION     V025841
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..3497
                     /label="ZNF717(NM_001128223)"
                     /note="ZNF717(NM_001128223)"
                     /gene="ZNF717"
     CDS             3504..3569
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3593..4181
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4247..4295
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4497..4925
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4939..5268
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5335..6129
                     /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    6305..6426
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6475..6491)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6499..6515
                     /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(6523..6553)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6568..6589
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6877..7465)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7636..8496)
                     /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(8497..8601)
                     /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 ccaccatgtt tccagtgttc tctggctgtt
      781 tccaagagct acaagaaaag aataaatctc tggagttggt gtcctttgag gaggtagctg
      841 tgcacttcac ctgggaggag tggcaggacc tggatgatgc tcagaggacc ctgtacaggg
      901 acgtgatgct ggagacctac agcagcctgg tatcattggg gcattacatt accaaacctg
      961 agatgatctt caagctagag caaggagcag agccatggat agtagaagaa accccaaacc
     1021 tgagactttc agctgtccag atcattgatg accttattga aaggagccat gaaagtcatg
     1081 atagattttt ctggcaaatt gtaatcacca acagcaacac atcaactcag gagagagttg
     1141 aattaggaaa aacatttaat ttgaactcaa accatgtttt aaatctgatt ataaataatg
     1201 gaaacagttc aggaatgaag cctgggcagt ttaatgattg ccagaacatg cttttcccta
     1261 ttaagcctgg ggagacacag tctggagaga aacctcatgt ctgtgatata accaggagat
     1321 cccacagaca tcatgaacat cttactcagc atcacaagat tcaaactctg ctgcagactt
     1381 ttcaatgtaa tgaacaaggg aaaaccttca acacggaggc aatgttcttt atacataaga
     1441 gggttcatat agtacagacc tttggtaaat ataatgaata tgagaaagcc tgtaataact
     1501 cagctgttat tgtccaagtg ataactcagg taggacagcc aacttgctgt agaaagtctg
     1561 acttcactaa acatcagcag acacacacag gagagaaacc ctatgaatgt gttgaatgtg
     1621 agaaaccctc cattagcaaa tcagacctta tgctacagtg caagatgcct actgaggaaa
     1681 aaccttatgc ctgtaactgg tgtgaaaaat tgttcagcta taagtccagc ctcattatcc
     1741 atcagagaat tcacacaggg gaaaagccct atggatgcaa tgaatgtggt aaaacctttc
     1801 gccgtaagtc attcctcact ttacatgaga gaactcacac aggggataaa ccctacaaat
     1861 gtattgaatg tggaaaaact tttcactgta agtcacttct cactttacat cacagaactc
     1921 actcagggga aaagccctat cagtgtagtg aatgtggaaa aacctttagc cagaagtcat
     1981 acctcacaat acatcataga actcacacag gggaaaagcc ctatgcatgt gaccattgtg
     2041 aagaagcatt tagccataag tcaaggctta ctgtccatca gagaacacac acaggggaaa
     2101 aaccgtatga atgtaatgag tgtggaaaac cctttatcaa taagtcaaac ctcaggttac
     2161 atcagagaac tcacacaggg gaaaaaccct atgaatgcaa tgaatgtggg aaaacgtttc
     2221 accgtaagtc attcctcact atccatcaat ggactcacac aggggaaaaa ccctacgaat
     2281 gcaatgaatg tgggaaaacc tttcgctgta agtcattcct cactgtccat cagagaactc
     2341 atgctgggga aaaaccatac gcatgtaacg aatgtggaaa aacatatagc cacaagtcat
     2401 accttacagt acatcacaga actcacacag gggaaaaacc ctatgaatgt aatgaatgtg
     2461 gaaaatcctt tcactgtaag tcattcctaa ctatacatca gagaactcat gctggcaaaa
     2521 aaccctatga atgtaatgaa tgtgagaaaa cctttatcaa taagttaaac cttgggatac
     2581 acaagagaac tcacacaggg gaaagaccct atgaatgtaa tgaatgtgga aaaacctttc
     2641 gtcagaagtc aaatctcagc acccatcagg gaactcacac aggagagaaa ccttacgtat
     2701 gtaatgaatg tggaaaaacc tttcatcgca agtcattcct caccatacac cagagaactc
     2761 acacggggaa aaaccgtatg gatgtaatga atgtggaaaa actttttgtc agaaatcata
     2821 ccttattata catcagagaa ctcacaccgg ggaaaagccc tatgaatgta atgaatgtgg
     2881 aaaatccttt catcagaagg caaatcttca gaagcatcaa ggtattcaca cgggggagaa
     2941 accctatgaa tgtagcaaat gtggaaaaac cttgtcagaa gtcagtcctc actgtacatc
     3001 atagaaccca taccggagaa aaaccttatg aatgtaatga gtgtgggaaa accttttgtc
     3061 acaagtcaaa cctcagtacg catcagggga ctcactcagg agagaaaccc tatgaatgtg
     3121 atgaatgtag gaaaactttt tacgataaga cagttctcac catacatcag agaactcaca
     3181 caggtgagaa gccatttgaa tgtaaagaat gtaggaaaac cttctcccag aagtcaaaac
     3241 tctttgtaca tcacagaact cacacagggg aaaaaccctt tagatgtaat gaatgtagga
     3301 aaactttctc ccagaagtca ggcctcagta tacatcagag aacacacaca ggagaaaaac
     3361 cttatgaatg taaggaatgt gggaaaacct tttgccagaa gtcacacctc agcaggcatc
     3421 aacaaaccca tataggagag aaatctgacg tagctgaggc aggctatgtg ttccctcaaa
     3481 atcactcttt tttccctgga tccgactaca aagaccatga cggtgattat aaagatcatg
     3541 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     3601 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3661 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3721 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3781 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3841 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3901 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3961 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4021 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4081 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4141 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4201 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4261 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4321 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4381 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4441 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     4501 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4561 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4621 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     4681 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4741 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4801 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4861 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4921 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4981 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5041 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5101 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5161 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5221 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5281 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5341 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5401 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5461 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5521 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5581 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5641 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5701 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5761 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5821 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5881 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5941 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6001 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6061 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6121 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6181 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6301 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6421 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8641 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     8701 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     8761 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
