LOCUS       V046207                 8739 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046207
VERSION     V046207
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..3419
                     /label="Zfp748(NM_001035231)"
                     /note="Zfp748(NM_001035231)"
                     /gene="Zfp748"
     CDS             3426..3491
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3515..4103
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4169..4217
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4419..4847
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4861..5190
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5257..6051
                     /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    6227..6348
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6397..6413)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6421..6437
                     /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(6445..6475)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6490..6511
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6799..7387)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7558..8418)
                     /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(8419..8523)
                     /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 ggaaatgctg tcattcaggg
      781 atgtggccgt tgatttctct gcagaggagt gggagtgcct ggagcctgct cagtggaatc
      841 tgtacaggga cgtgatgctg gagaattaca gccaccttgt gttcctgggt cttgctttct
      901 ctaagccata cctggtcacg tttctggagc aaagacaaga gcctttgaat gagaagagac
      961 aagtggccac caccatgcac ccagatataa aaccatataa atgtaaagaa tgtggcaaag
     1021 cctgtgattg gaactcactc cttctcgaac accagagaac caatccagga gagaatgcct
     1081 acaagtgtga agaatgtggt caggcttctg gctcttactc agtaattcct gaacaccata
     1141 taaacgatac ggaagagaaa gcctgcaagt gtgaagaatg tggtaaagtc atttgtactt
     1201 gctcagaaaa ttctagctac cagagaattt gcattggaga aaacccctac aagtgtgaag
     1261 aatgtggaaa agccttttct tctcattcat gtcttgctca acatgaggta gaacacactg
     1321 gacagcaatt ttataactgt gaagaatgtg ggaaactgtt ttattgtcct tctcacctta
     1381 cagaacatca aaaaattcat tctcaagaga atctctttaa gattgaggta tgtagcgagg
     1441 tcttttgtgc tcccatagaa ctttctaaag accagaactt ttgtactgaa gaaaaaccct
     1501 acaggtatga agactatgtt aaggccttta gcgcttgctc agttctttct gaacacccga
     1561 caattcatcc aggagagaaa gcattcaagt gtgaagaatg tggcaatgcc ttttgtacac
     1621 tccattcagt ctctaaaatg aaaattcatt atgaagtgaa atcctacaag tgtgaagagt
     1681 gtgggaaagc ctttgctact catctttccc tgattcagca taagataggt catactcgag
     1741 agaaacctta tcaatgtgaa gaatgtggca aaatgtttta ctgttcttca aaccttaagc
     1801 aacatcaaat aactcattct caagagaaac cctacaagtg tgaagtatgt ggcaaggtct
     1861 ttagaacttg ctggcaactt tctaaacacc tgagaatcca ttctggagag aaaccataca
     1921 agtgtgagga atgtggcaaa gccttttata ctctctccta ccttactcag cacaaattag
     1981 gtcacactgg ggagaaacct tacaaatgtg aggaatgtgg caaaaccttt tactaccctt
     2041 cagtccttaa ggaacaccta gcaattcatt ctggagagaa accgtacagg tgtgatgaat
     2101 gtggcaagga cttttgtact cgctcaggac gttccagaca ccagagaatc catactggag
     2161 agaaaccgta caagtgtgag caatgtggaa aggccttcag tacccattca tacctttctc
     2221 atcacaagat agttcacact ggacataaac cctacaagtg tgaagagtgt ggcaaaaaat
     2281 tttattatcc ttctcgactt aaggagcatc aaaggattca ttctcaagag aatccctaca
     2341 agtgtgaaat atgtggcaaa gccttttata ctcactcata ctttacccag cacaaattag
     2401 gtcacactgg ggagaaacct tacaaatgtg aggaatgtgg caaaaccttt tactaccctt
     2461 caatccttaa ggaacaccta gcaattcatt ctggaaagaa accgtacagg tgtgaagaat
     2521 gtggcaagga cttttgtact cgctcaggac gttccagaca ccaaagaatc catactggag
     2581 agaaacccca caagtgtgaa gaatgcggaa aagtcttttc tactcattca tatcttactc
     2641 agcacaaagt agtccactct ggagagaagc cctacaagtg tgaagagtgt ggcaaaaagt
     2701 tttactatcc ttctcgcctt aaggagcatc aaaggattca ttctcaagag aatccttaca
     2761 agtgtgaaat atgtggcaat gtattttgta ctcccaaagg actttcgaaa caccagagat
     2821 ttcatactgg agagaaaccc tacaagtgtg aagaatgcgg gaaaatgttt tattatcctt
     2881 ctcgacttaa ggaacatcaa agaattcatt ctcaagagaa tccttacaag tgtgaaatat
     2941 gtggcaaagc cttttatact cactcctacc ttactcagca caaattaggt catactgggg
     3001 agaaacctta caaatgtgag gaatgtggca aaacctttta ctacccttca atccttaagg
     3061 aacacctagc aattcattct ggaaagaaac cgtacaggtg tgaagaatgt ggcaaggact
     3121 tttgtactcg ctcaggacgt tccagacacc agagaatcca tactggagag aaaccctaca
     3181 agtgtgagca atgtggaaag gccttcagta ctcactcata cctttctcaa cacaaggtag
     3241 tccactctgg agagaagccc tacaagtgtg aagagtgtgg caaaatgttt tactattctt
     3301 ctcgacttaa ggagcatcaa aggattcatt ctcaagagaa tccctacaag tgtgaagtgt
     3361 gtggcaaagt ctttagtgct cacttagaac ttgctaccca cctaagtatt cattccggag
     3421 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3481 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3541 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3601 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3661 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3721 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3781 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3841 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3901 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3961 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4021 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4081 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4141 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4201 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4261 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4321 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4381 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4441 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4501 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4561 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4621 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4681 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4741 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4861 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4921 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4981 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5041 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5101 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5161 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5221 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5281 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5341 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5401 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5461 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5521 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5581 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5641 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5701 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5761 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5821 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5881 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5941 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6001 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6061 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6121 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6181 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6241 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6301 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6361 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6421 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6481 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6541 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6601 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6661 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6781 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6841 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6901 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6961 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7021 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7081 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7141 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7201 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7261 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7321 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7381 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7441 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7501 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7561 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7621 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7681 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7741 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7801 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7861 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7921 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7981 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8041 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8101 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8161 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8221 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8281 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8341 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8401 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8461 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8521 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8581 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8641 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8701 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
