LOCUS       V031000                 5889 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031000
VERSION     V031000
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..2036
                     /label="Rnf2(NM_001360845)"
                     /note="Rnf2(NM_001360845)"
                     /gene="Rnf2"
     CDS             2055..2084
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2103..2126
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2146..2162)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2173..2795
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2944..3532)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3861..3908
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4140..4934)
                     /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(4969..5326)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5328..5432)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5459..5887
                     /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 gtctcaggct gtgcagacaa atggaactca accattaagc aaaacatggg
     1081 aactcagttt gtatgagtta caacgaacac ctcaggaggc aataacagat ggcttggaaa
     1141 ttgtggtttc acctagaagt ctacacagtg aattaatgtg cccaatttgt ttggatatgt
     1201 taaagaacac catgactaca aaggagtgtt tacatcggtt ttgcgcggat tgtattatca
     1261 cagcccttag aagtggcaac aaagagtgtc ctacctgtcg gaaaaaactg gtttctaaaa
     1321 gatcactaag gccagacccg aactttgatg cactcatcag caagatttat cccagtcgtg
     1381 atgagtatga agcgcatcag gaaagggtct tagcaaggat caacaaacac aacaatcagc
     1441 aggctctcag ccacagcatc gaggaggggc tgaagataca ggccatgaac agattacagc
     1501 gaggcaaaaa gcagcagata gaaaatggta gtggagcaga agataatggt gacagctccc
     1561 actgtagtaa cgcatccaca cacagcaacc aggaagcggg cccgagtaac aaacggacca
     1621 aaacctctga tgactctggg cttgaacttg ataacaacaa tgcagcagtg gcgattgatc
     1681 cagtcatgga cggtgccagt gagattgagt tagtcttcag gccccatcca actcttatgg
     1741 aaaaggacga cagcgcacag acaagataca taaagacttc aggcaatgcc actgttgatc
     1801 acttatccaa gtatctggct gtgaggttag ctttagaaga acttcgaagc aaaggagaat
     1861 caaaccagat gaacctggat acagccagtg agaagcagta caccatttac atagccacag
     1921 ccagtggcca gttcaccgtt ttaaatggct ccttttcttt ggaattggtc agtgagaaat
     1981 actggaaagt gaacaaaccc atggaacttt attatgcacc caccaaggag cacaaaacgc
     2041 gtacgcggcc gctcgagcag aaactcatct cagaagagga tctggcagca aatgatatcc
     2101 tggattacaa ggatgacgac gataaggttt aaacggccgg ccgcggtcat agctgtttcc
     2161 tgaacagatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     2221 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     2281 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2341 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     2401 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     2461 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     2521 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2581 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     2641 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     2701 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     2761 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ggcgagcggt
     2821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3661 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
     3721 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     3781 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     3841 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     3901 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     3961 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     4021 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     4081 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     4141 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     4201 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     4261 gtagccaacg ctatgtcctg atagcgatcc gccacaccca gccggccaca gtcgatgaat
     4321 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     4381 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     4441 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     4501 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     4561 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     4621 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     4681 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     4741 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     4801 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     4861 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     4921 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     4981 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5041 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5101 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5161 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5221 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5281 acaccctaac tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt
     5341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5581 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5821 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5881 gcgaatttt
//
