LOCUS       V027187                 8413 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027187
VERSION     V027187
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3906
                     /label="Tmem67(NM_177861)"
                     /note="Tmem67(NM_177861)"
                     /gene="Tmem67"
     CDS             3913..3939
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3980..4204
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4250..4678
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4692..5021
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5088..5882
                     /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    6056..6177
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6226..6242)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6250..6266
                     /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(6274..6304)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6319..6340
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6628..7213)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7384..8244)
                     /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(8245..8349)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggtgacgc gtacaaggcc tgtggcggcc
      961 atggcggtca gatctcgttc ttcatcccgg actggcaccg cctaccttct cttagttctc
     1021 tgcgaagtct catgggcaca aatcttctcc ttccctttcc ggcggcctga gacctgcgac
     1081 ttcaaccagt actttgacat ctcggcgctc tcctgtgccc cctgtggcgc caaccagcgg
     1141 cgggatgcac taggaacttc ctgtgtatgt ctaccaggat atcatatgat ctctaataat
     1201 ggaggacctt ctattatttg taaaaagtgt ccagaaaaca tgaaaggcgt tactaaagat
     1261 ggctgggact gcatttcttg tcctagtggc ttaactgctg aggggaagtg ccactgtccc
     1321 actggccata tattagtgga aagaaatgtt agtgggtctt tgttagccca agcaacttgc
     1381 gagctttgtg atgaaagtga aaactcattt acaaaagcca atgctttagg aaccaggtgt
     1441 gtccgctgcg aaccaacatt tgtgaataca agcaggtcct gttcatgttc agagcctcac
     1501 actttaacag gtggattatg tttcagcaac acagggaatt ttcatcaacg tgtaatttcc
     1561 actgcacgtt atggagagct tggcatgtca ttaaattcag aatggtttgc caagtatttg
     1621 caggcaacag cagctgcatg ctggactcac gccaacctga catcgtgcca agctcttgga
     1681 aatatgtgcg tgatgaacat gaactcttac gactcgacca cactcgatgc ttgtcgactg
     1741 ttccattaca tctttgaaag cactgctgga ttgatcagtg tgcattctgt tccattctgg
     1801 agacagaacc ttccttggct gttctatgga gaccagccag gattggctcc tcaagttctc
     1861 agtactacac ctcttcctac aaatttcagt tttaaaggac aaaaccagct gaagtttgtt
     1921 gctgcttcct atgatatacg aggaaacttc atcaaatggc aacctttaga aggtggtgtt
     1981 ttacagcttt gtccagacac tgagagaagg ctggatgcag cgtatgcttt tggaacaacg
     2041 taccagcaaa attgtgaaat ctctctctct aaactcctgg ttgacttcag cagccctgta
     2101 ttttatgatg tttacctcga atacactgat gaagagcagc accgttacct ttggcctatc
     2161 ccagtgttaa acctaaatct tcaacacaac aaactgtttg tgaaccaaga cagtagctcc
     2221 agcaagtggc ttctgactcg gcgcattttc ttagtggatg cagtaagtgg gcgagaaaat
     2281 gacctaggaa atcagccaag agtaattcga gttgctaccc agatttcact gagcatacgc
     2341 cttgtaccca acacaaaaaa tggaaacata tacactcccc tacttaccat tgcctacagt
     2401 gacattgaca tcaaaaatgc tcatagccag tctgcgaaga tttccttctc agtgaagtat
     2461 gagatgaatc aaggagacgc atctgtgcac acagatattg cgttgggtgt gttgggtggg
     2521 ctggctgtgc tatcatctct tttgaagaca gcgggatgga agaggcgcat cgggagcccc
     2581 atgattgatt tacagacagt tatgaaattc ctgctgtact atgctggtga tctggccaat
     2641 gtgtttttta tcatcacagt gggaacaggt ctttattggc ttattttctt caaagcacag
     2701 aagtcagtct ctgttttgct gccgatgcca gttcaggaag aacgtttcgt tacttatgtg
     2761 gggtgcgcct ttgctatgaa ggctctccag tttttacata agttcatatc ccagatttcc
     2821 atagatatat tctttattga ttgggaacgg ccgaaaggaa aagttcttaa agctgttgaa
     2881 ggtgagggtg gtgtgcggag tgccactgtt cctgtgagca tatggagaac gtatttcgtg
     2941 gcaaatgaat ggaatgagat tcagactgtg agaaaaatta atccactctt tcaagtgctt
     3001 accacactct tcttcttgga ggttgtggga ttcaaaaatt tagcgttaat ggattcatcc
     3061 tccagccttt ctagaaaccc atcagactac acagctcctt acagccgcat tctgagatat
     3121 gccgtggcta ctgctatctg gctggtgatt ggaataatac aggttgtttt ctttgctgcc
     3181 ttttatgaga gattcataga agacaaaatc cggcagtttg ttgacctgtg ctccatgagt
     3241 aatgtgtcag tgttcctgct gtcccacaga tgttttggat attacattca tgggaggtca
     3301 gtccacggcc atgcagacac taacatggag gaaatgaata tgaaccttaa gagagaggcg
     3361 gaaaatttgt gcagccaaag aggtttggtg cccaacacgg atgggcagac gtttcagatt
     3421 gcagtttcta gtcagatgag acaacactat gacaggattc atgagacttt aacaaggaga
     3481 aatggccctg ccagactgct gagttcgtcg gggagtactt ttgaacaaag catcaaagca
     3541 taccacgcaa tgaataagtt ccttggctct ttcattgacc acgttcataa ggaaatggat
     3601 tactttataa aagataagtt gcttcttgaa agaatccttg ggatggaatt catggagccc
     3661 atggagaaga gcatcttcta taatgatgaa ggccattctt tcagcagtgt gctgtattat
     3721 gggaacgagg ccacgctcct catctttgat ctgttgttct tctgtgttgt ggatttggct
     3781 tgccaggact ttgttttagc atccttcctc acatacctgc aacaagagat tttccgattt
     3841 atccgaaata cagtaggaca aaagaatttg gcaaccaaaa cactagtgga tgaaagattt
     3901 ctgatcctcg agtacccata cgacgtacca gattacgctt agtctagagg gcccgtttaa
     3961 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4021 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4081 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4141 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4201 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4261 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4321 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4381 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4441 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4501 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4561 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4621 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4681 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4741 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4801 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4861 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4921 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4981 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5041 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5101 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5161 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5221 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5281 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5341 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5401 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5461 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5521 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5581 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5641 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5701 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5761 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5821 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5881 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5941 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6001 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6061 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6121 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6181 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6301 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6421 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7141 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8401 gatcccctat ggt
//
