LOCUS       V030067                 7077 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030067
VERSION     V030067
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..3224
                     /label="Kcnn3(NM_080466)"
                     /note="Kcnn3(NM_080466)"
                     /gene="Kcnn3"
     CDS             3243..3272
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3291..3314
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3334..3350)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3361..3983
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4132..4720)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5049..5096
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5328..6122)
                     /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(6157..6514)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6516..6620)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6647..7075
                     /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 ggacacttct gggcacttcc atgactcggg ggtgggggac ctggatgaag
     1081 accccaagtg tccctgtcca tcttctgggg acgagcaaca gcagcagcag caaccgccac
     1141 caccgccagc gccaccagca gtcccccagc agcctccggg acccttgctg cagcctcagc
     1201 ctccgcagcc tcagcagcaa cagtcgcagc agcagcagca gcagcagtcg cagcagcagc
     1261 agcagcaggc tccactgcac cccctgcctc agcttgccca actccagagc cagcttgtcc
     1321 atcctggtct gttgcactct tctcccacgg cgttcagggc ccccacttca gccaactcta
     1381 ccgccatcct ccacccttcc tccaggcaag gcagccagct aaatctcaat gaccacttgc
     1441 ttggccactc tccaagttcc acagccacaa gtgggcccgg tggaggcagc cggcaccggc
     1501 aggccagccc cctggtgcac cggcgggaca gcaatccctt cacggagata gctatgagct
     1561 cctgcaaata cagcggtggg gtcatgaagc ccctcagccg cctcagcgcc tctcggagaa
     1621 accttatcga ggctgagcct gagggccaac ccctccagct cttcagtccc agcaaccccc
     1681 cagagattat catctcctcc agggaggata accatgccca ccagactctg ctccatcacc
     1741 ccaacgctac ccacaaccac cagcatgccg gcaccacggc cggcagcacc accttcccca
     1801 aagccaacaa gcggaaaaac caaaacattg gctataagct gggacacagg agggccctgt
     1861 ttgaaaagag aaagcgactg agtgactatg ctctgatttt tgggatgttt ggaattgttg
     1921 ttatggtgat agagaccgaa ctgtcttggg gtttgtactc aaaggattcc atgttttcgt
     1981 tggccctgaa atgccttatc agtttatcca ccgtcatcct gcttggtttg atcatcgcct
     2041 atcacacaag ggaagtacag ctctttgtga tcgacaatgg tgcagatgac tggcggatag
     2101 ccatgaccta cgagcgtatc ctctacatca gcctggagat gctggtgtgc gccatccacc
     2161 ccatccctgg agagtacaag ttcttctgga ctgcacgcct ggccttctcc tataccccat
     2221 ctcgggcaga ggctgacgtg gacattattc tgtccatccc catgttcctg cgcctgtacc
     2281 tgatcgcccg ggtcatgctg ctccatagca aactcttcac tgatgcctcc tcccgaagca
     2341 tcggggccct caacaagatc aacttcaaca cccgattcgt catgaagacg ctcatgacca
     2401 tctgccctgg cacggtgctg ctggtgttca gcatctctct gtggatcatc gctgcctgga
     2461 ctgtgcgagt ctgtgaaagg taccatgatc agcaggatgt aactagtaac tttctgggtg
     2521 ccatgtggct catctccatc acgttccttt ccattggcta tggggacatg gtgccccaca
     2581 catactgtgg gaaaggtgtc tgtcttctca ccggcatcat gggtgcaggc tgcactgccc
     2641 ttgtggtagc tgtggttgcc cgaaagctcg aactcaccaa agcagagaag catgtgcaca
     2701 acttcatgat ggacactcag ctcaccaaac ggatcaagaa tgccgccgcc aatgtcctcc
     2761 gggaaacgtg gctaatctat aaacatacaa agctgctaaa gaagattgac catgccaaag
     2821 tcaggaaaca ccagaggaag ttcctccaag ctatccacca actgcggggt gtcaagatgg
     2881 aacaaaggaa gctgagtgac caagccaaca ccctggtgga cctttccaag atgcagaacg
     2941 tcatgtatga cttaatcacg gagctcaatg accggagtga agatctagaa aagcagattg
     3001 gcagcctgga gtccaagctg gagcacctca cagccagttt caattccctg cccctgctca
     3061 tcgcagatac cctgcgccag cagcagcagc agctgctcac tgccttcgtg gaggcccggg
     3121 gcatcagtgt ggccgtggga actagccacg cccctccctc tgacagccct atcgggatca
     3181 gctccacctc tttcccaacc ccatacacaa gttcaagcag ttgcacgcgt acgcggccgc
     3241 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     3301 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     3361 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3421 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3481 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3541 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3601 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3661 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3721 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3781 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3841 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     3901 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     3961 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc
     4021 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4861 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     4921 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     4981 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     5041 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     5101 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     5161 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     5221 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     5281 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     5341 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     5401 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     5461 atgtcctgat agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     5521 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     5581 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     5641 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     5701 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     5761 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     5821 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     5881 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     5941 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     6001 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     6061 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     6121 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     6181 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     6241 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     6301 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     6361 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     6421 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     6481 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     6661 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6721 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6781 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6841 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6901 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6961 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7021 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttt
//
