LOCUS       V041707                 7663 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041707
VERSION     V041707
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..3102
                     /label="GRAMD1A(NM_020895)"
                     /note="GRAMD1A(NM_020895)"
                     /gene="GRAMD1A"
     CDS             3109..3135
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3139..3165
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3169..3195
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3230..3454
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3500..3928
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3942..4271
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4338..5132
                     /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    5306..5427
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5476..5492)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5500..5516
                     /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(5524..5554)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5569..5590
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5878..6463)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6634..7494)
                     /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(7495..7599)
                     /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 atgttcgaca ccacacccca ctctggccgg
      961 agcacgccaa gcagctcccc atcgctccgg aaacggctgc agctcctgcc cccaagccgg
     1021 cccccacctg agccagaacc aggcaccatg gtggagaagg gatcagatag ctcctcagag
     1081 aagggtgggg tgcctgggac ccccagcacc cagagcctag gcagccggaa cttcatccgc
     1141 aacagcaaga agatgcagag ctggtacagt atgctgagcc ccacttataa gcagcgtaat
     1201 gaggacttcc ggaaactgtt cagcaaactc cccgaagcag aacgcctcat tgtggattac
     1261 tcctgcgccc tgcagcgtga gatcctgctc cagggccgcc tctacctctc tgagaactgg
     1321 atctgcttct acagcaacat cttccgctgg gagaccacga tctccatcca gctgaaggaa
     1381 gtgacatgtc tgaagaagga aaagacggcc aagctgatcc ccaacgccat ccagatctgc
     1441 acggagagcg agaagcattt cttcacttcc tttggggccc gtgaccgctg cttcctcctc
     1501 atcttccgcc tctggcagaa tgcactgctt gaaaagacgc tgagtccccg cgagctctgg
     1561 cacctggtgc atcagtgcta cggctcagag ctgggcctca ccagtgagga tgaggactat
     1621 gtctccccct tgcagctgaa cggtctgggg acccccaagg aagtgggaga tgtgatcgcc
     1681 ctgagcgaca tcacctcctc gggggcagct gaccgcagcc aggagccaag cccagtgggt
     1741 tcgcgccgtg gccatgtcac gcccaacctt tcccgagcca gcagcgacgc agaccatggg
     1801 gcagaggagg acaaggagga gcaggtagac agccagccag acgcctcctc cagccagaca
     1861 gtgaccccgg tggctgaacc cccgagcaca gagcccaccc agcctgacgg gcccaccacc
     1921 ctgggcccct tggatctgct gcccagtgag gagctattga cagacacaag taactcctct
     1981 tcatccactg gggaggaagc ggacttggct gccctgcttc ccgacctctc cggccgcctc
     2041 ctcatcaact ctgtcttcca tgtgggcgct gagcggctcc agcagatgct cttctcggac
     2101 tcgcccttcc tccagggctt cctacagcag tgcaagttca cagacgtgac gctgagcccc
     2161 tggagtgggg acagcaagtg ccaccagcgc cgggtgctga cgtacaccat ccccatcagc
     2221 aacccactgg gccccaagag cgcctccgtg gtggagacac agacgctgtt ccggcgcggc
     2281 ccccaggccg gcgggtgtgt ggtggactcc gaggtgctga cgcagggcat cccctaccag
     2341 gactacttct acactgccca ccgctactgc atcctgggtc tggcccggaa caaggcgcgg
     2401 ctccgagtgt cttctgagat ccgctaccga aagcagccgt ggagcctggt gaagtcgctc
     2461 attgagaaga actcgtggag cggcattgaa gactatttcc accatctgga gcgagagctc
     2521 gccaaggctg agaagctgtc tctggaggaa ggcgggaagg atgcccgggg cttgctatcc
     2581 ggcctgcggc ggcggaagcg gcccctgagc tggcgggctc acggggacgg gccccagcac
     2641 ccagatcctg acccctgtgc ccgggccggc attcacacct cgggctccct cagctcccgc
     2701 ttctccgaac catctgtgga ccagggcccc ggggcaggca tccccagtgc cctggttctc
     2761 atcagcattg tgatctgtgt gagccttatc atcctcatcg ccctcaacgt cctgctcttc
     2821 taccgcctct ggtccctgga aaggacagcc cacacctttg agtcctggca cagcctggcc
     2881 ctggccaagg gcaagttccc ccagacggcc acagagtggg ccgagatcct ggcgctgcag
     2941 aagcaattcc acagcgtgga ggtgcacaag tggaggcaga tcctgcgggc ctccgtggag
     3001 ctcctggatg agatgaagtt ctcgctggag aagctgcacc aaggcatcac agtctcagac
     3061 cctccctttg acacccagcc ccggcccgat gacagctttt ccctcgagta cccatacgac
     3121 gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta tccttatgat
     3181 gtcccagact acgcatgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3241 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3301 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3361 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3421 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3481 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3901 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4201 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4261 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4321 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4381 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4441 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4501 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4561 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4621 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4681 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4741 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4801 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4861 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4921 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4981 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5041 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5101 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5161 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5221 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5281 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5341 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5401 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5461 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5521 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5581 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5641 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5701 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5761 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5821 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5881 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5941 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6001 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6061 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6121 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6181 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6241 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6301 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6361 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7621 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
