LOCUS       V043595                 8757 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043595
VERSION     V043595
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..3437
                     /label="Zbtb21(NM_001081685)"
                     /note="Zbtb21(NM_001081685)"
                     /gene="Zbtb21"
     CDS             3444..3509
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3533..4121
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4187..4235
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4437..4865
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4879..5208
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5275..6069
                     /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    6245..6366
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6415..6431)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6439..6455
                     /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(6463..6493)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6508..6529
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6817..7405)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7576..8436)
                     /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(8437..8541)
                     /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 ccaccatgat gcacagtgta cagcgggctc
      781 acaaacactc gggagaggaa gagagaagaa agatgtgtgc ttgccagata aagtgcagga
      841 tggagggact gctgcattac atcaacccag cacatgctat ctctctgctc agtgccctca
      901 atgaggagcg cctcaaggga cagctgtgtg acgtgctcct gattgttggg gaccagaaat
      961 tccgagctca taagaacgtc ttggctgcca gcagtgagta cttccagagt ttattcacga
     1021 ataaggagaa cgaggcacag actgtctttc agctggactt ctgtgagcct gacgcttttg
     1081 acaacgttct gaactacatt tactcttctt ccctttttgt ggagaagggc agccttgctg
     1141 ctgtgcagga gctggggtat agccttggta tctccttcct gaccaacatc gttgccaaag
     1201 cccctcaggc tccttttcca gcctgtccca acaggaaaag agtgccagtg gaagatgatg
     1261 aaaccagctc tcaaaagcga agtgtcattg tgtgtcaggg cagaagtgaa gtgccaggga
     1321 aagccagtgg tccagccatg caggacctca gccatactgc ccgggcctcc cccagtgctt
     1381 cagtcaagac caccatcagt aagccacatg tggccaagcc gccagagcag cttcatagtc
     1441 tgcccttaac tgaaaagagc tggacaaagg atagtgctgc agtatatgca aagtctctgg
     1501 aacaggctgg ggctttggat gatcctaata ggggcagttt ggtaaagaga aatacagtcc
     1561 tgcccccaaa gccttcacag gacagggagg ccacagatga taaaccaggg gtgagcagcc
     1621 agcttcccaa ggggaaagct atagagctgg ctctgaagag accacggccc cctgttctgt
     1681 ctcttcatag ctcatcagag actccatatc tcctaaaaga aactagcaaa ggaggcggtc
     1741 agggagagga taggaacttg ctctactact ctaagctagg gctggtggtc ccttctggtg
     1801 ggcctgcttc tgcaaaccag agcattgaca gaagtggccc actagtgaaa agcctcctca
     1861 ggcggtcact gtctatggac agccaggttc ctgtttactc cccctccata gatttgaagt
     1921 catcccaggg atcatccaca gcggcaaatg aggcaccggg tagtgtgttc tgttcgatgt
     1981 ctcaaaagtc atctttaaaa gattgcagtg aaaaaaaagc tctggatgac aggcctcaag
     2041 tgctccagcc tcatcgcctc aggtccttta gtgcttccca gtcaacagac agggaggaag
     2101 cctcccctgt gactgaggta cgcattaaga cggaacccag cagcccactg tcagacccct
     2161 cagacatcat ccgggtcact gtgggaggag acgcagcagc agcagcagta gcagcagcag
     2221 cagcagcagc agctacgaga gacctgcccc tcaaaacaga ggaagaccag agagatatga
     2281 gcagactccc agcaaagaga aggttccaga cagacagaag gtcacccttg aagaaggcaa
     2341 aggcaaatga acacgggcct gctgtctcag aagagaactg tgaagagggc aggagccctc
     2401 cttcccttga cagcaacttc ccagattctg acttaaacag agaggagttt gagcaaggca
     2461 accacgagcg cttgtgccgg aacgccaccg tttgccctta ctgcagcctc aggttcttct
     2521 ctcccgcact gaagcaggaa catgaggaca ggtgtgagta caaaaagctg acttgcctgg
     2581 agtgcatgcg caccttcaag tcctccttca gcatctggcg gcaccaggta gaagtgcata
     2641 accagaacaa catggcttta gcagagaact tcgccctggc caccctggac cacaacggtg
     2701 aagtggcagc cgcttccagg cctcagactg agcccagcaa ggtaaaccat gtggccactc
     2761 caaaagagga tacagcattt agcgactctt cagagcaagt gaactttgat tctgaggatt
     2821 cctcctgcct ccctgaagac ttgagtcttt caaagcaact gaaagtccaa gtcaaagagg
     2881 agcctgtgga ggaggcagag gaggaggctc ctgaggctag tgcagctccc agggaggctg
     2941 gccccagcaa ggagaccggc ctatggccct gtgagaaatg cggaaagatg ttcacggcac
     3001 acaagcagct ggagcggcac caggagctgc tgtgttctgt gaagcccttc atctgccacg
     3061 tatgctacaa agccttccgt accaacttcc gactctggag tcacttccag acccacatgt
     3121 ctcaggctac agaggagcct gtgcagaaag aatcagaggt gtgtcctgtg cccacaaact
     3181 ccccctcccc accacctctg ccacctccgc cacccttgcc caagattcag cccctggagc
     3241 cagacagccc cacaggcctg cctgagaacc caaccccagc tacagagaag ctgtttgcac
     3301 cccaggagtc agataccctc ttctaccacg ccccacccct ttcagcaatc acatttaaaa
     3361 gacagttcat gtgcaagctc tgccatagga cattcaagac ggccttcagt ctttggagtc
     3421 atgagcagac acacaatgga tccgactaca aagaccatga cggtgattat aaagatcatg
     3481 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     3541 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3601 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3661 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3721 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3781 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3841 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3901 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3961 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4021 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4081 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4141 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4201 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4261 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4321 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4381 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     4441 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4501 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4561 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     4621 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4681 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4741 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4801 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4861 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4921 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4981 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5041 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5101 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5161 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5221 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5281 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5341 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5401 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5461 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5521 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5581 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5641 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5701 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5761 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5821 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5881 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5941 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6001 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6061 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6121 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6181 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6241 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6361 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8581 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     8641 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     8701 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
