LOCUS       V045172                 8799 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045172
VERSION     V045172
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..3479
                     /label="Zfp598(NM_183149)"
                     /note="Zfp598(NM_183149)"
                     /gene="Zfp598"
     CDS             3486..3551
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3575..4163
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4229..4277
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4479..4907
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4921..5250
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5317..6111
                     /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    6287..6408
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6457..6473)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6481..6497
                     /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(6505..6535)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6550..6571
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6859..7447)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7618..8478)
                     /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(8479..8583)
                     /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 ggcggcggcg ggcgccgagg
      781 ggcggcgcgc ggctctggag gcggtggcag cgccggagcg gggcggcggg agctgcgtgc
      841 tgtgctgcgg tgacctggag gctacggcgc tgggccgctg cgatcacccg gtgtgctacc
      901 ggtgctctac caagatgcgg gtgctgtgcg agcagcgcta ctgcgccgtg tgccgcgagg
      961 agctgcgcca ggtggtcttt gggaagaaac tccctgcttt tgccctgatc cccatccatc
     1021 agctccaaca cgagaagaaa tatgacatct attttgcaga tggaaaggtg tttgcattgt
     1081 acaggcagct gctgcagcat gagtgcccac ggtgccctca cctgccaccc ttcagcctct
     1141 tcggggactt ggagcagcac atgcgcaagc agcatgagct cttctgctgc aagttgtgcc
     1201 tcaagcacct caagatcttc acctatgaac gcaagtggta ctcacgcaag gacctggctc
     1261 gacaccgtat gcagggtgac cctgatgata catcccatcg gggacatcca ctctgtaagt
     1321 tctgtgacga gcgctacctg gacaatgacg agctgctcaa gcacctgcgc cgagaccact
     1381 acttctgcca cttctgtgac tctgatgggg cccaggacta ctatagtgac tatgcctacc
     1441 tgcgtgagca cttccgggag aagcacttcc tgtgtgagga gggccgctgc agcactgagc
     1501 agttcaccca tgccttccgc accgagatcg acctcaaggc tcataaaaca gcttgccaca
     1561 gccgcagccg ggccgaggca cgccagaacc ggcagattga ccttcagttt agctttgcac
     1621 cacggcactc acgccggagt gagggagtcg tcagtggtga ggactacgag gaggtggaca
     1681 ggtacaatcg ccagggccga gctggccggg ccagcgggcg tggagcccag cagaaccgcc
     1741 ggggaagctg gaggtacaag agggaagaag aagaccgaga agtggcagct gctatccggg
     1801 cctctgtggc tgctcaacag caagaggaga ctcagagggt tgaggaccgg gaggagggta
     1861 gcaggcccaa gaaagaggaa gccgcagcca gggtgcctga ggaaccccgt ggccaccggc
     1921 gtctgcctcg ggctcagggt gaaggctcag gctccaagga agcctcagca aatggtcctg
     1981 taagccaaga agccttccca gccacaggcc caggcccagt ggttgccctc tcaaataccc
     2041 tcccaccacc cagccctgag ctcaaggagg aagacttccc cagtctctgt gcctccacct
     2101 cctcctgctg cacagcggtt accccaggct ctgtaggctt ggcactggct tatcctggtc
     2161 ctcccagggg caagaacacc ttccaggaag aggactttcc tgccttggtt tcctccgcac
     2221 ccaagcccag cagtgcccct tccagtctca tctcagcttg gaatagcggc tgcagcaaga
     2281 aagggaacct acccaccccg gggtcccagg ctgtggttgg tggcagccag cctccccgga
     2341 aggcaggaaa ggggagcagg ggtggcagaa agggtggccc agccccagtg gatgaggagg
     2401 atagcggtgg cctcactgtg caggggctgc ggagtgtgcc caccacggta gctgtgtctt
     2461 ctttgctggc accagccacc aaccagagtt cagccaaggt tggcaagaag aagaaggtgg
     2521 gctctgagaa gcctggggcc acatcatccc cactgttgcc ccctgatcat accccaaagc
     2581 cctctggggc tgagcaggtc ctggaagccc ctttgagcaa agctgaagtg ccagtaacca
     2641 tcgttgtcaa tggacattcc gaagggtcag ccctggtacg gagtgcccca aaggaacctc
     2701 cagggcttcc aaggcccctc ggtcccctcc cctgccccat accacaggag gactttccag
     2761 ctctcggagg cccttgtcca cccaggatgc ctcctccccc aggcttcagc accgtggtac
     2821 tcttgaaggg cacaccacct ccacccccac ccccacccgg cctggtgcct cccatcagca
     2881 agccaccccc tggcttctct agtcttctgc ccagctccca ctcagcctgt gctcctagtc
     2941 ccaccaccac aaccaccacc accaccacca cgaaaacacc cgggctagca cccacaccac
     3001 aagcctactt agttcctgag aatttccggg agagaaacct gcagcttatc cagtctatca
     3061 aggactttct gcaaagcgat gaggcctgct tcagcaagtt taaaagccat tcgggggagt
     3121 tcagacaggg gatgatctct gcggctcagt actacaagag ctgcagggac ctgcttggag
     3181 agagcttcca gaagatcttc agtgagctgc tggcgctcct gccagacaca gccaaacagc
     3241 aggagctgct gtctgcacac acagacttct gcagtcgtga gaagcctccc aactccagat
     3301 ctaagaggaa caagaagaat gtgtggcaga ccagcaccca acagttgggc ctggactgct
     3361 gtgtgtgccc cacctgccag caggtactgg cacacggtga cgtcagcagc caccaggcac
     3421 tgcacgctgc ccgggacgat gacttcccct cccttcaagc cattgccagg atcattacgg
     3481 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3541 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3601 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3661 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3721 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3781 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3841 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3901 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3961 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4021 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4081 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4141 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4201 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4261 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4321 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4381 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4441 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4501 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4561 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4621 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4681 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4741 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4801 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4861 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4921 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4981 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5041 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5101 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5161 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5221 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5281 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5341 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5401 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5461 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5521 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5581 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5641 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5701 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5761 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5821 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5881 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5941 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6001 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6061 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6121 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6181 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6241 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6421 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6481 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7021 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7261 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7501 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7561 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7621 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7681 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7741 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7801 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7861 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7921 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7981 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8041 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8101 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8161 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8221 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8281 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8341 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8401 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8461 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8581 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8641 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8701 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8761 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
