LOCUS       V020601                 7823 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020601
VERSION     V020601
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"
     CDS             872..2503
                     /label="Zbtb7b(NM_001355206)"
                     /note="Zbtb7b(NM_001355206)"
                     /gene="Zbtb7b"
     CDS             2510..2575
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2599..3187
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3253..3301
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3503..3931
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3945..4274
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4341..5135
                     /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    5311..5432
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5481..5497)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5505..5521
                     /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(5529..5559)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5574..5595
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5883..6471)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6642..7502)
                     /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(7503..7607)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgttaca gaaggaaact acaaggggcc agctgtggtg
      721 tccctcagtc tggagcccct gcaccaggat cgggggactt ggtgggagct taacccccac
      781 ggcaccggga agcaaccagc tcccctcgcc tccttccccc ttcgtggctt gcggtctctc
      841 ttccccgcct cggcccccag gaagtgagaa gatggggagc cccgaggatg acctgattgg
      901 aatcccattc ccagaccaca gcagcgagct cctgagctgc ctgaacgagc agcgccagct
      961 gggccatctg tgcgacctca ccatcaggac gcagggtctt gagtaccgca cccacagggc
     1021 tgtgctggct gcctgtagcc actacttcaa gaagctcttt acggagggag gagggggaac
     1081 cgtgatggga acagggggtg gtgggacggc ttctggggga gcaggggccg gtgtgtgtga
     1141 gctggacttt gtagggccag aggccctagg tgccctgctc gagtttgctt acacagccac
     1201 actgaccacc agcagcgcca atatgccggc tgtactccaa gctgctcggc tactggaaat
     1261 cccgtgtgtc atcgctgctt gcatggagat tctacaaggc agtgggctgg aagcccccag
     1321 ccccgatgag gatgactgtg agcgagcccg acagtacctg gaggcttttg ccactgccac
     1381 caccacagcc tcaacctcag gaatgcccaa cggtgaagac agccctccac aggtgcccct
     1441 cctaccgccg ccgccaccgc cacctcgacc tgttgcccgt cgcagccgca aaccccggaa
     1501 agcttttctt caaaccaaag gggcccgagc aaaccatctg gtccctgagg cacccacagt
     1561 actcacccat cccttgacct acgaagaaga agagatggtt ggtagattgg gtaacagtgg
     1621 gggcagtggg ctcggagata gctatagtcc tcctacaggt gccgcctcac ctgccgaggg
     1681 gcccctgaac tatgaagtct ttgaaggtga ggaagaagag gaggagatgg catacccccc
     1741 aggctacggg ctagcccaga gtaatgagcc ctcgctatca ccagaggagc tgggctcaga
     1801 tgaagatccc atcgaccccg acctgatggc ttacctaagt tcgctgcacc aggacgccct
     1861 gacaccaggc ctggatggcc aagacaagct ggtgcgtaaa cgccgttcac agatgcccca
     1921 agagtgccca gtctgtcaca agataatcca cggggcaggc aaactgcctc ggcacatgag
     1981 gacccacact ggtgagaagc cctttgcctg tgaggtctgc ggcgtccgct tcaccaggaa
     2041 tgacaagctg aagatccaca tgcggaagca cacaggagaa cgcccctact cgtgccccca
     2101 ctgcccagcc cgcttcctgc atagctacga cctcaagaac cacatgcacc tgcacaccgg
     2161 ggaccggccc tatgagtgcc acctgtgcca caaggccttc gccaaggagg accacctgca
     2221 gcgccatctc aagggtcaga actgcctgga ggtgcgcacc cgaaggcgcc gcaaggatga
     2281 cgtggcagcc cctcactacc caccgccctc gaccaccacc tcatcccctg ctggcctcga
     2341 cctttccaat ggccacctgg acaccttcca cctctctctg gctcgattct gggagcagtc
     2401 ggccaccacg ggacccccag tcactacaca agggccccct gaggaggagg aagaggaagg
     2461 gacacccacc acaccacagg ccgaaggtgc catggagtcc tctctcgagg actacaaaga
     2521 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     2581 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2641 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2701 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2761 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2821 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2881 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2941 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3001 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3061 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3121 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3181 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3241 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3301 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3361 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3421 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3481 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     3661 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3841 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3901 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4141 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4201 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4261 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4321 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4381 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4441 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4501 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4561 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4621 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4681 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4741 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4801 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4861 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4921 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4981 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5041 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5101 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5161 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5221 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5281 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5341 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5401 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5461 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5521 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5581 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5641 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5701 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5761 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5821 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5881 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5941 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6001 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6061 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6121 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6181 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6241 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6301 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6361 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6421 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6481 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6541 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6601 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6661 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6721 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6781 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6841 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6901 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6961 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7021 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7081 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7141 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7201 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7261 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7321 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7381 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7441 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7501 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7561 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7621 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     7681 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7741 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7801 attgcatgaa gaatctgctt agg
//
