LOCUS       V031018                 5337 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031018
VERSION     V031018
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..1484
                     /label="Ube2b(NM_001362685)"
                     /note="Ube2b(NM_001362685)"
                     /gene="Ube2b"
     CDS             1503..1532
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1551..1574
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(1594..1610)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    1621..2243
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2392..2980)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3309..3356
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(3588..4382)
                     /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(4417..4774)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4776..4880)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      4907..5335
                     /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 gtcgaccccg gcccgtagga ggctcatgcg ggatttcaag cgattgcaag
     1081 aggacccacc tgtgggggtc agtggcgccc catctgaaaa caacatcatg cagtggaatg
     1141 cagttatatt tggaccagaa gggacaccct ttgaagatgg tacttttaaa ctagtaatag
     1201 aattttctga agaatatcca aataaaccac caaccgttag gtttttatcc aaaatgtttc
     1261 atccaaatgt gtatgctgat ggtagcatat gtttagatat cctgcagaat cgatggagtc
     1321 ccacatatga tgtctcttcc atcttaactt ccattcagtc tctgctggat gaaccgaatc
     1381 caaacagtcc agccaacagc caagcagcac agctttatca ggaaaacaaa cgggagtatg
     1441 agaagagagt ttcggccatt gttgagcaaa gctggaatga ttcaacgcgt acgcggccgc
     1501 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     1561 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     1621 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     1681 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     1741 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     1801 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     1861 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     1921 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     1981 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     2041 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     2101 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     2161 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     2221 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc
     2281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3121 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     3181 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     3241 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     3301 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     3361 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     3421 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     3481 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     3541 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     3601 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     3661 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     3721 atgtcctgat agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     3781 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     3841 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     3901 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     3961 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     4021 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     4081 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     4141 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     4201 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     4261 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     4321 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     4381 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     4441 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     4501 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     4561 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     4621 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     4681 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     4741 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     4801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     4921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4981 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5041 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5101 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5221 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5281 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttt
//
