LOCUS       V044993                 9786 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044993
VERSION     V044993
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..4466
                     /label="Zfat(NM_001145888)"
                     /note="Zfat(NM_001145888)"
                     /gene="Zfat"
     CDS             4473..4538
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4562..5150
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5216..5264
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5466..5894
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5908..6237
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6304..7098
                     /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    7274..7395
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7444..7460)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7468..7484
                     /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(7492..7522)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7537..7558
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7846..8434)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8605..9465)
                     /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(9466..9570)
                     /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 gacgcggacg gcagaaaaca
      781 cagccatctt catgtgtaaa tgctgtaatc ttttctcacc aaatcagtca gaactcgtga
      841 cccacgtttc tgagaagcat ggggaagaag gggtcaatgt ggatgacgtc atcattcccc
      901 tccggcctct gaacactcca gaaaacccca acccaagcaa gggtggagat gagtttttgg
      961 ttatgaagag aaagagaggg aggcctaagg gttccacaaa gaagcccagc actgaagagg
     1021 aggtggtaga aaacctcgtg agccccagtg aggacggtcc cctggctaca gaggaaggga
     1081 gcagactggc ccccagcagc ttggagtgta gcaagtgctg ccggaagttc tccaacacac
     1141 gccagctgcg caagcacatc tgcatcattg tgctgaatct gggtgaggag gatggagatg
     1201 caggtaacga gtcagacctt gatctagaaa agacatacaa agaagatgat cgagaaaagg
     1261 cttcaaaaag accccgggct cagaaaacag agaaagtcca gaagatctca ggaaaggagg
     1321 cagggcagct ttctggagcg aagaaaccca tcatcagcgt ggtcttaact gctcatgaag
     1381 caattccagg tgctaccaag attatcccag tggaggctgg gcctcccgaa acaggagcac
     1441 cccctccaga gaccactgca gcagacctgg tgccgcggag agggtaccaa gaatacgcca
     1501 tccagcagac cccatatgag cagcccatga agtcaagcag gttaggtccc acccagctga
     1561 agatcttcac ttgtgagtac tgcaacaagg tgttcaagtt caagcactct ctgcaggccc
     1621 acctgaggat ccacacgaac gagaagccat acaagtgctc ccagtgcagc tatgccagcg
     1681 ccatcaaggc caacctcaac gttcacctgc gcaagcacac cggagagaaa ttcgcctgcg
     1741 actactgctc cttcacctgc ctcagcaagg gccacctcaa ggtgcacatc gagcgggtac
     1801 acaagaagat caaacagcat tgccgcttct gcaagaagaa atactcagat gtcaagaacc
     1861 tcatcaagca catccgggac atgcatgacc cgcaggacaa gaaggtcaag gaggccttag
     1921 acgagctccg cctcatgaca agggagggca agcggcagct gctctacgac tgccacatct
     1981 gcgagcgcaa gttcaagaat gagctagacc gagaccgcca catgctggtt catggtgaca
     2041 agtggccctt tgcctgcgag ctctgtggcc atggggccac taagtaccag gccctggaac
     2101 tgcatgtccg aaagcacccc tttgtgtatg tctgtgctct atgtctcaag aagttcgtca
     2161 gctccatcag gctgcgctcc catatccgag aggtgcatgg ggcggcccag gagaccttgg
     2221 tgtttaccag ctccatcaac cagagtttct gcctcctgga gccgggcggg gacatccagc
     2281 aggaagcctt gggggaccag ctacagctgg cagctgagga atttgtgtgt ccagaaattg
     2341 atgtacgcaa ggaggaggct tgtcctgggg aagctcagcc tgaggtgggg ttgagggagt
     2401 tggtggtccc tggagatgca catgcccctc cacctgggcc cttggccacc ccccagagtg
     2461 aaagtagttc cctgtccccc tgcaaactag aaaccactgt ggtctcctcc gacctcaact
     2521 ctcttggagt ggtttcagat gattttttac tgaaaagcga tacctcctct gctgagcctc
     2581 ccgctgctgc cgaggcaacc tcggatacac agcatcgaga ctcagcccag actcagggtg
     2641 aagaagtcac actgctgctg gccaaggcca aaagtgctgg accagaccca gagagctctc
     2701 cgggtggaag gcagaaggtg ggtgctctgc cagccagtga atctgactct agcacatgtc
     2761 tcagggcaaa cccaacagaa gcctcagacc tccttcctac agtgactgat ggaggggacc
     2821 ttggtgtgtg ccagcctgac tcctgcacac catcctctga gcaccaccct ggcagcacag
     2881 cattcatgaa ggtcctggac agtctccaga agaagcagat gaacaccagt ctttgtgagc
     2941 ggatccggaa ggtttacgga gacctggagt gtgaatactg cggcaaactt ttttggtacc
     3001 aggtgcattt tgacatgcat gtccgcaccc acacccggga acatctgtat tattgctccc
     3061 agtgtcacta ttcttccatc accaaaaact gccttaaacg ccatgtaatt cagaaacaca
     3121 gtaacatctt gctgaagtgc cccactgacg gctgtgacta ctcgactcca gataaatata
     3181 agctgcaggc ccatcttaaa gttcacacag agctggacaa aaggagttat tcttgtcctg
     3241 tatgtgaaaa atcattttca gaagaccgat tgataaagtc acatatcaag actaatcatc
     3301 cagaggtctc catgaatacc atttctgagg ttcttgggag aagagtccag ctcaaagggc
     3361 taattggaaa gcgagccatg aagtgtccat attgtgattt ctatttcatg aagaatggct
     3421 cagaccttca gcggcacatc tgggctcatg agggtgtgaa gcccttcaag tgttctctgt
     3481 gtgaatatgc aactcgtagc aagagcaacc tcaaagctca tatgaatcgt cacagcactg
     3541 agaagaccca cctctgtgac atgtgtggca agaaattcaa atccaaaggg acactgaaaa
     3601 gtcataagct ccttcacacg tctgatggga agcaattcaa gtgcacggtg tgtgactaca
     3661 cagcagccca gaaaccccag ctgctgcgac acatggagca gcatgcctcc ttcaagcctt
     3721 tccgctgtgc tcactgtcat tactcttgta acatctctgg atctctgaaa agacactaca
     3781 acaggaagca ccccaatgag gagtacacca acgtgggcag cggggagctc gcagctgaag
     3841 ccctcatcca gcaaggtggt ctgaagtgtc ctgtttgcag ctttgtgtat ggaaccaaat
     3901 gggagttcaa cagacacttg aagaacaaac atggcttgaa ggtagtggag attgatggag
     3961 accccaaatg ggagccagca gcagagactc ccgaggagcc ctccacccag tatctctaca
     4021 tcactgaagc tgaagatgtt caggggacac aagcagctgt ggctgcactt caggacctga
     4081 gatatacctc cgagagtggt gatcgacttg accccaccgc tgtgaatatc ctgcagcaga
     4141 tcattgaact aggttcggag acccacgatg ctgctgctgt ggcctctgtg gttgccatgg
     4201 cgcccggcac agtgactgtt gtaaagcagg tcaccgatga ggagcccagt tccaaccaca
     4261 cagtcatgat tcaggagact ctccagcagg cctccgtgga gctggccgag cagcaccatc
     4321 tggtggtgtc ctctgacgac gtggagggca tagagacagt gacggtgtac acacagggtg
     4381 gggaggcctc agagttcatc gtatacgtgc aagaggctgt acagcccgtg gaggagcagg
     4441 ttggggagca gccagcccca gaactcggat ccgactacaa agaccatgac ggtgattata
     4501 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     4561 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4621 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4681 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4741 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4801 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4861 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4921 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4981 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5041 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5101 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     5161 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5221 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     5281 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5341 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5401 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     5461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     5641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5821 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     6001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     6181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     6301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7081 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     7141 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7201 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7261 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7321 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7381 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7441 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7501 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7561 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7621 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7801 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8041 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8281 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8581 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8641 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8701 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8761 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8821 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8881 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8941 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9001 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9061 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9121 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9181 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9241 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9301 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9361 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9421 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9481 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9541 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9601 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     9661 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9721 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9781 cttagg
//
