LOCUS       V046749                 7755 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046749
VERSION     V046749
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..2435
                     /label="ZNF250(NM_021061)"
                     /note="ZNF250(NM_021061)"
                     /gene="ZNF250"
     CDS             2442..2507
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2531..3119
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3185..3233
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3435..3863
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3877..4206
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4273..5067
                     /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    5243..5364
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5413..5429)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5437..5453
                     /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(5461..5491)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5506..5527
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5815..6403)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6574..7434)
                     /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(7435..7539)
                     /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 agcagccaga ctcctgccag
      781 tgccggcagg accccagccc ctgtcgttcc aggccaagct gaccttcgag gatgtggctg
      841 tgctcctctc ccaggatgaa tgggaccgcc tgtgccctgc tcagaggggt ctctacagaa
      901 atgtgatgat ggaaacctat gggaatgtag tctcattggg acttccagga tccaagcctg
      961 acataatctc ccagctggag cgaggggaag atccctgggt cctggacagg aagggggcta
     1021 agaagagcca gggcctgtgg agtgactact cagacaacct caaatatgac cacactacag
     1081 cctgtacaca acaagacagt ttatcttgtc catgggaatg tgaaaccaag ggagagagtc
     1141 aaaatacaga cttgagtccg aagccattaa tttcagagca aacagtgatt ctggggaaaa
     1201 cacccttggg gaggattgat caagaaaata atgaaacaaa gcaaagcttc tgtctgagtc
     1261 caaactctgt tgaccaccgt gaagttcagg tcttaagcca aagcatgcca ctcactccgc
     1321 accaggcagt gcctagtgga gagaggccct acatgtgtgt tgagtgtggg aagtgctttg
     1381 gccggagttc ccacctcctt cagcatcagc gtatccacac tggagagaag ccctatgtgt
     1441 gcagtgtatg tgggaaggcc ttcagccaga gctcagtcct tagtaaacac aggagaattc
     1501 acacaggtga gaagccctat gagtgtaatg agtgtggaaa agcctttaga gtgagctcag
     1561 atcttgctca gcatcacaag atacatacag gagagaagcc tcacgaatgt cttgagtgtc
     1621 ggaaagcctt cactcaactc tcacatctca ttcagcacca gcggatccac acgggagaaa
     1681 ggccatatgt gtgtccgttg tgtgggaaag ccttcaacca tagcactgtt ctgcggagcc
     1741 accagagggt acacactggg gagaagcctc acaggtgcaa tgagtgtggg aaaaccttca
     1801 gtgtgaagag gacactgctg cagcaccaga ggatccacac cggggagaag ccctacacgt
     1861 gcagcgagtg tgggaaggcc ttcagcgacc gctcagtcct cattcagcac cacaacgtgc
     1921 acaccgggga gaagccctat gagtgcagtg agtgtgggaa gaccttcagc caccgctcca
     1981 cactgatgaa tcacgagcgg atccacaccg aggaaaagcc ctatgcatgc tacgaatgtg
     2041 ggaaggcctt cgttcagcac tcacacctga tccagcacca gagagtccac actggggaga
     2101 agccctatgt gtgtggtgaa tgtgggcacg ccttcagtgc acgccggtct ctgatccagc
     2161 atgagagaat ccacacaggt gaaaagccct tccagtgcac agaatgtggc aaagccttca
     2221 gcctgaaagc aactctgatt gtgcacctga ggacccacac gggcgagaag ccatatgagt
     2281 gcaatagctg cgggaaggcc ttcagccagt actcagtgct catccagcac cagcggatcc
     2341 acacaggcga gaagccctat gagtgcgggg agtgtgggcg tgccttcaac cagcatggcc
     2401 acctaatcca gcaccagaaa gtgcacagaa agttgggatc cgactacaaa gaccatgacg
     2461 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     2521 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2941 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3121 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3181 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3241 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3301 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3361 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3421 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3481 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3541 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3601 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3661 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3721 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3781 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3841 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3901 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3961 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4081 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4141 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4201 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4261 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4321 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4381 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4441 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4501 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4561 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4621 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4681 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4741 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4801 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4861 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4921 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4981 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5041 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5101 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5161 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5221 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5341 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5401 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5461 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5521 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6001 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6241 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6541 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6601 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6661 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6721 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6781 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6841 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6901 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6961 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7021 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7081 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7141 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7201 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7261 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7321 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7381 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7561 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7621 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7681 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7741 aagaatctgc ttagg
//
