LOCUS       V018216                10887 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018216
VERSION     V018216
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1029..7034
                     /label="TET2(NM_001127208)"
                     /note="TET2(NM_001127208)"
                     /gene="TET2"
     CDS             7053..7082
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             7101..7124
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(7144..7160)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    7171..7793
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(7942..8530)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    8859..8906
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(9138..9932)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(9967..10324)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(10326..10430)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      10457..10885
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggaacaggat agaaccaacc atgttgaggg caacagacta agtccattcc
     1081 tgataccatc acctcccatt tgccagacag aacctctggc tacaaagctc cagaatggaa
     1141 gcccactgcc tgagagagct catccagaag taaatggaga caccaagtgg cactctttca
     1201 aaagttatta tggaataccc tgtatgaagg gaagccagaa tagtcgtgtg agtcctgact
     1261 ttacacaaga aagtagaggg tattccaagt gtttgcaaaa tggaggaata aaacgcacag
     1321 ttagtgaacc ttctctctct gggctccttc agatcaagaa attgaaacaa gaccaaaagg
     1381 ctaatggaga aagacgtaac ttcggggtaa gccaagaaag aaatccaggt gaaagcagtc
     1441 aaccaaatgt ctccgatttg agtgataaga aagaatctgt gagttctgta gcccaagaaa
     1501 atgcagttaa agatttcacc agtttttcaa cacataactg cagtgggcct gaaaatccag
     1561 agcttcagat tctgaatgag caggagggga aaagtgctaa ttaccatgac aagaacattg
     1621 tattacttaa aaacaaggca gtgctaatgc ctaatggtgc tacagtttct gcctcttccg
     1681 tggaacacac acatggtgaa ctcctggaaa aaacactgtc tcaatattat ccagattgtg
     1741 tttccattgc ggtgcagaaa accacatctc acataaatgc cattaacagt caggctacta
     1801 atgagttgtc ctgtgagatc actcacccat cgcatacctc agggcagatc aattccgcac
     1861 agacctctaa ctctgagctg cctccaaagc cagctgcagt ggtgagtgag gcctgtgatg
     1921 ctgatgatgc tgataatgcc agtaaactag ctgcaatgct aaatacctgt tcctttcaga
     1981 aaccagaaca actacaacaa caaaaatcag tttttgagat atgcccatct cctgcagaaa
     2041 ataacatcca gggaaccaca aagctagcgt ctggtgaaga attctgttca ggttccagca
     2101 gcaatttgca agctcctggt ggcagctctg aacggtattt aaaacaaaat gaaatgaatg
     2161 gtgcttactt caagcaaagc tcagtgttca ctaaggattc cttttctgcc actaccacac
     2221 caccaccacc atcacaattg cttctttctc cccctcctcc tcttccacag gttcctcagc
     2281 ttccttcaga aggaaaaagc actctgaatg gtggagtttt agaagaacac caccactacc
     2341 ccaaccaaag taacacaaca cttttaaggg aagtgaaaat agagggtaaa cctgaggcac
     2401 caccttccca gagtcctaat ccatctacac atgtatgcag cccttctccg atgctttctg
     2461 aaaggcctca gaataattgt gtgaacagga atgacataca gactgcaggg acaatgactg
     2521 ttccattgtg ttctgagaaa acaagaccaa tgtcagaaca cctcaagcat aacccaccaa
     2581 tttttggtag cagtggagag ctacaggaca actgccagca gttgatgaga aacaaagagc
     2641 aagagattct gaagggtcga gacaaggagc aaacacgaga tcttgtgccc ccaacacagc
     2701 actatctgaa accaggatgg attgaattga aggcccctcg ttttcaccaa gcggaatccc
     2761 atctaaaacg taatgaggca tcactgccat caattcttca gtatcaaccc aatctctcca
     2821 atcaaatgac ctccaaacaa tacactggaa attccaacat gcctgggggg ctcccaaggc
     2881 aagcttacac ccagaaaaca acacagctgg agcacaagtc acaaatgtac caagttgaaa
     2941 tgaatcaagg gcagtcccaa ggtacagtgg accaacatct ccagttccaa aaaccctcac
     3001 accaggtgca cttctccaaa acagaccatt taccaaaagc tcatgtgcag tcactgtgtg
     3061 gcactagatt tcattttcaa caaagagcag attcccaaac tgaaaaactt atgtccccag
     3121 tgttgaaaca gcacttgaat caacaggctt cagagactga gccattttca aactcacacc
     3181 ttttgcaaca taagcctcat aaacaggcag cacaaacaca accatcccag agttcacatc
     3241 tccctcaaaa ccagcaacag cagcaaaaat tacaaataaa gaataaagag gaaatactcc
     3301 agacttttcc tcacccccaa agcaacaatg atcagcaaag agaaggatca ttctttggcc
     3361 agactaaagt ggaagaatgt tttcatggtg aaaatcagta ttcaaaatca agcgagttcg
     3421 agactcataa tgtccaaatg ggactggagg aagtacagaa tataaatcgt agaaattccc
     3481 cttatagtca gaccatgaaa tcaagtgcat gcaaaataca ggtttcttgt tcaaacaata
     3541 cacacctagt ttcagagaat aaagaacaga ctacacatcc tgaacttttt gcaggaaaca
     3601 agacccaaaa cttgcatcac atgcaatatt ttccaaataa tgtgatccca aagcaagatc
     3661 ttcttcacag gtgctttcaa gaacaggagc agaagtcaca acaagcttca gttctacagg
     3721 gatataaaaa tagaaaccaa gatatgtctg gtcaacaagc tgcgcaactt gctcagcaaa
     3781 ggtacttgat acataaccat gcaaatgttt ttcctgtgcc tgaccaggga ggaagtcaca
     3841 ctcagacccc tccccagaag gacactcaaa agcatgctgc tctaaggtgg catctcttac
     3901 agaagcaaga acagcagcaa acacagcaac cccaaactga gtcttgccat agtcagatgc
     3961 acaggccaat taaggtggaa cctggatgca agccacatgc ctgtatgcac acagcaccac
     4021 cagaaaacaa aacatggaaa aaggtaacta agcaagagaa tccacctgca agctgtgata
     4081 atgtgcagca aaagagcatc attgagacca tggagcagca tctgaagcag tttcacgcca
     4141 agtcgttatt tgaccataag gctcttactc tcaaatcaca gaagcaagta aaagttgaaa
     4201 tgtcagggcc agtcacagtt ttgactagac aaaccactgc tgcagaactt gatagccaca
     4261 ccccagcttt agagcagcaa acaacttctt cagaaaagac accaaccaaa agaacagctg
     4321 cttctgttct caataatttt atagagtcac cttccaaatt actagatact cctataaaaa
     4381 atttattgga tacacctgtc aagactcaat atgatttccc atcttgcaga tgtgtagagc
     4441 aaattattga aaaagatgaa ggtccttttt atacccatct aggagcaggt cctaatgtgg
     4501 cagctattag agaaatcatg gaagaaaggt ttggacagaa gggtaaagct attaggattg
     4561 aaagagtcat ctatactggt aaagaaggca aaagttctca gggatgtcct attgctaagt
     4621 gggtggttcg cagaagcagc agtgaagaga agctactgtg tttggtgcgg gagcgagctg
     4681 gccacacctg tgaggctgca gtgattgtga ttctcatcct ggtgtgggaa ggaatcccgc
     4741 tgtctctggc tgacaaactc tactcggagc ttaccgagac gctgaggaaa tacggcacgc
     4801 tcaccaatcg ccggtgtgcc ttgaatgaag agagaacttg cgcctgtcag gggctggatc
     4861 cagaaacctg tggtgcctcc ttctcttttg gttgttcatg gagcatgtac tacaatggat
     4921 gtaagtttgc cagaagcaag atcccaagga agtttaagct gcttggggat gacccaaaag
     4981 aggaagagaa actggagtct catttgcaaa acctgtccac tcttatggca ccaacatata
     5041 agaaacttgc acctgatgca tataataatc agattgaata tgaacacaga gcaccagagt
     5101 gccgtctggg tctgaaggaa ggccgtccat tctcaggggt cactgcatgt ttggacttct
     5161 gtgctcatgc ccacagagac ttgcacaaca tgcagaatgg cagcacattg gtatgcactc
     5221 tcactagaga agacaatcga gaatttggag gaaaacctga ggatgagcag cttcacgttc
     5281 tgcctttata caaagtctct gacgtggatg agtttgggag tgtggaagct caggaggaga
     5341 aaaaacggag tggtgccatt caggtactga gttcttttcg gcgaaaagtc aggatgttag
     5401 cagagccagt caagacttgc cgacaaagga aactagaagc caagaaagct gcagctgaaa
     5461 agctttcctc cctggagaac agctcaaata aaaatgaaaa ggaaaagtca gccccatcac
     5521 gtacaaaaca aactgaaaac gcaagccagg ctaaacagtt ggcagaactt ttgcgacttt
     5581 caggaccagt catgcagcag tcccagcagc cccagcctct acagaagcag ccaccacagc
     5641 cccagcagca gcagagaccc cagcagcagc agccacatca ccctcagaca gagtctgtca
     5701 actcttattc tgcttctgga tccaccaatc catacatgag acggcccaat ccagttagtc
     5761 cttatccaaa ctcttcacac acttcagata tctatggaag caccagccct atgaacttct
     5821 attccacctc atctcaagct gcaggttcat atttgaattc ttctaatccc atgaaccctt
     5881 accctgggct tttgaatcag aatacccaat atccatcata tcaatgcaat ggaaacctat
     5941 cagtggacaa ctgctcccca tatctgggtt cctattctcc ccagtctcag ccgatggatc
     6001 tgtataggta tccaagccaa gaccctctgt ctaagctcag tctaccaccc atccatacac
     6061 tttaccagcc aaggtttgga aatagccaga gttttacatc taaatactta ggttatggaa
     6121 accaaaatat gcagggagat ggtttcagca gttgtaccat tagaccaaat gtacatcatg
     6181 tagggaaatt gcctccttat cccactcatg agatggatgg ccacttcatg ggagccacct
     6241 ctagattacc acccaatctg agcaatccaa acatggacta taaaaatggt gaacatcatt
     6301 caccttctca cataatccat aactacagtg cagctccggg catgttcaac agctctcttc
     6361 atgccctgca tctccaaaac aaggagaatg acatgctttc ccacacagct aatgggttat
     6421 caaagatgct tccagctctt aaccatgata gaactgcttg tgtccaagga ggcttacaca
     6481 aattaagtga tgctaatggt caggaaaagc agccattggc actagtccag ggtgtggctt
     6541 ctggtgcaga ggacaacgat gaggtctggt cagacagcga gcagagcttt ctggatcctg
     6601 acattggggg agtggccgtg gctccaactc atgggtcaat tctcattgag tgtgcaaagc
     6661 gtgagctgca tgccacaacc cctttaaaga atcccaatag gaatcacccc accaggatct
     6721 ccctcgtctt ttaccagcat aagagcatga atgagccaaa acatggcttg gctctttggg
     6781 aagccaaaat ggctgaaaaa gcccgtgaga aagaggaaga gtgtgaaaag tatggcccag
     6841 actatgtgcc tcagaaatcc catggcaaaa aagtgaaacg ggagcctgct gagccacatg
     6901 aaacttcaga gcccacttac ctgcgtttca tcaagtctct tgccgaaagg accatgtccg
     6961 tgaccacaga ctccacagta actacatctc catatgcctt cactcgggtc acagggcctt
     7021 acaacagata tataacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
     7081 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc
     7141 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg
     7201 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     7261 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     7321 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     7381 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     7441 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     7501 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     7561 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     7621 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     7681 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     7741 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg
     7801 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7861 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8341 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     8401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8461 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     8701 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     8761 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     8821 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     8881 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     8941 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     9001 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     9061 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     9121 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     9181 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     9241 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc
     9301 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     9361 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     9421 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     9481 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     9541 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     9601 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     9661 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     9721 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     9781 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     9841 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     9901 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     9961 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
    10021 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
    10081 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
    10141 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
    10201 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
    10261 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
    10321 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    10381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    10441 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
    10501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
    10561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
    10621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
    10681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
    10741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
    10801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
    10861 atttaacaaa aatttaacgc gaatttt
//
