LOCUS       V021421                 8123 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021421
VERSION     V021421
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             683..2803
                     /label="ZCCHC8(NM_017612)"
                     /note="ZCCHC8(NM_017612)"
                     /gene="ZCCHC8"
     CDS             2810..2875
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2899..3487
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3553..3601
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3803..4231
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4245..4574
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4641..5435
                     /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    5611..5732
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5781..5797)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5805..5821
                     /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(5829..5859)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5874..5895
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6183..6771)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6942..7802)
                     /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(7803..7907)
                     /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 ccatggccgc agaggtgtat tttggcgatc tagagctctt
      721 cgagccgttc gaccacccag aggagtcgat tccgaagccc gttcacactc gcttcaagga
      781 cgacgacggc gacgaggagg acgaaaatgg ggtcggcgac gcggagctac gggagcggct
      841 tcggcagtgc gaggagacca tcgagcagct ccgcgccgag aatcaagaac ttaaacgaaa
      901 attgaacatt ctgactcgac cgagtggaat attggtgaac gatactaagt tagatggacc
      961 tatattacag attctattca tgaacaatgc tatttcaaag caatatcatc aagaaataga
     1021 ggaatttgta tcaaatttag taaaaagatt tgaggaacag cagaaaaatg atgtggaaaa
     1081 gacttccttt aatcttttgc cccagccatc cagtattgtg ctagaggagg accacaaagt
     1141 ggaagagtcc tgtgccatta aaaacaacaa ggaagctttc agtgttgtag gaagtgtcct
     1201 gtattttact aatttttgcc ttgataaatt ggggcaaccg cttctaaatg aaaaccctca
     1261 gctttccgaa ggatgggaaa tacccaagta ccatcaagtc ttcagccaca ttgtttctct
     1321 agaagggcaa gaaatacaag taaaggcaaa aaggccaaag cctcactgtt tcaattgtgg
     1381 ttctgaagaa caccaaatga aagattgccc aatgcctcgg aatgctgctc gaataagtga
     1441 aaagagaaaa gagtatatgg atgcctgtgg agaagcaaac aatcagaatt tccagcagcg
     1501 ataccacgca gaagaagtag aagaaagatt tggaagattc aagccaggag ttattagtga
     1561 ggaacttcaa gatgcactag gtgtgacaga caagagtctt ccacctttta tctatcggat
     1621 gcgccagcta gggtacccac cagggtggct caaagaggct gaattggaga attcggggct
     1681 tgcactctat gatggaaaag atggcactga tggggaaaca gaagttggag aaatacaaca
     1741 gaataaaagt gtcacttacg atctctcaaa attggtcaac tatcctggtt ttaatatatc
     1801 tactcccaga ggaattccag acgaatggag gatctttggt tccataccaa tgcaggcatg
     1861 tcagcagaag gatgtgtttg ccaattacct tacttctaac ttccaagcgc caggtgtgaa
     1921 gtctggcaac aagaggtctt catctcactc tagcccaggt agtccaaaga agcagaagaa
     1981 tgaaagcaac tcagcgggat ctcccgccga catggagctc gattcagata tggaggtacc
     2041 acatggttct cagagcagcg aaagttttca gtttcaacca ccattacctc ctgacactcc
     2101 tccactcccc cggggaactc ctccacccgt cttcacccct ccactcccaa agggcacccc
     2161 gccgctgact cccagtgact caccccagac cagaacagca tctggagctg tggatgagga
     2221 cgcactgact ctagaagaac ttgaagaaca gcagaggcgg atctgggcag ctcttgagca
     2281 ggccgagagc gtaaacagcg actccgacgt tcctgtggac acacctttaa ctggcaattc
     2341 cgttgcctca tcaccttgtc caaatgagct agacctccct gtcccggagg gaaaaacatc
     2401 tgaaaagcag acgctggatg agcctgaggt accagagatt tttacaaaga aatcagaagc
     2461 tggacatgcc tccagtccag actctgaggt gacatcactt tgtcagaagg aaaaagcaga
     2521 gttggctccg gtaaacactg aaggtgccct tcttgataat ggcagtgtcg taccaaactg
     2581 tgacatcagc aatgggggca gccagaagct ctttcctgca gacaccagtc cttcaacggc
     2641 cactaaaatt catagcccta tacctgacat gagcaaattt gcaactggaa tcacgccatt
     2701 tgaatttgag aatatggcag aatctactgg aatgtacctc aggataagaa gcttgttaaa
     2761 gaactcaccc cgaaaccagc agaaaaacaa aaaggcctct gaactcgagg actacaaaga
     2821 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     2881 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2941 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3001 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3061 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3121 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3181 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3301 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3361 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3421 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3481 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3541 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3601 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3661 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3721 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3781 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3841 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     3961 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4141 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4201 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4261 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4321 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4381 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4441 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4501 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4561 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4621 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4681 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4741 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4801 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4861 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4921 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4981 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5041 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5101 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5161 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5221 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5281 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5341 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5401 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5461 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5521 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5581 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5701 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5761 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5821 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5881 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5941 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6001 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6061 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6361 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7921 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     7981 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8041 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     8101 attgcatgaa gaatctgctt agg
//
