LOCUS       V023563                 7360 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023563
VERSION     V023563
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..2019
                     /label="GCDH(NM_000159)"
                     /note="GCDH(NM_000159)"
                     /gene="GCDH"
     CDS             2026..2052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2056..2082
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2086..2112
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2136..2724
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2790..2838
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3040..3468
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3482..3811
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3878..4672
                     /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    4848..4969
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5018..5034)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5042..5058
                     /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(5066..5096)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5111..5132
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5420..6008)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6179..7039)
                     /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(7040..7144)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc cctgagaggc
      721 gtctccgtgc ggctgctgag ccgcggaccc ggcctgcacg tccttcgcac gtgggtctcg
      781 tcggcggcgc agaccgagaa aggcgggaga acacagagcc aactggctaa gtcctcgcgt
      841 cccgagtttg actggcagga cccgctggtg ctggaggagc agctgaccac agatgagatc
      901 ctcatcaggg acaccttccg cacctactgc caggagagac tcatgcctcg catcctgttg
      961 gccaatcgca acgaagtttt tcatcgggag atcatttcgg agatggggga gttgggtgtg
     1021 ctgggcccca ccatcaaagg atatggctgt gctggggttt cgtctgtggc ctatgggctc
     1081 ctggcccgag agctggagcg ggtggacagt ggctacaggt cggcgatgag tgtccagtcc
     1141 tccctcgtca tgcaccctat ctatgcctat ggcagcgagg aacagcggca gaagtacctg
     1201 ccccagctgg ccaaggggga gctcctgggc tgcttcgggc tcacagagcc caacagcgga
     1261 agtgacccca gcagcatgga gaccagagcc cactacaact catccaacaa gagctacacc
     1321 ctcaatggga ccaagacctg gatcacgaac tcgcctatgg ccgatctgtt tgtagtgtgg
     1381 gctcggtgtg aagatggctg cattcggggc ttcctgctgg agaaggggat gcggggtctc
     1441 tcggccccca ggatccaggg caagttctcg ctgcgggcct cagccacagg catgatcatc
     1501 atggacggtg tggaggtgcc agaggagaat gtgctccctg gtgcatccag cctggggggt
     1561 cccttcggct gcctgaacaa cgcccggtac ggcatcgcgt ggggcgtgct tggagcttcg
     1621 gagttctgct tgcacacagc ccggcagtac gccctcgaca ggatgcagtt tggtgtccca
     1681 ctggccagga accagctgat tcagaagaag ctggcagaca tgctcactga gattaccctg
     1741 ggccttcacg cctgcctgca gctcggccgc ttgaaggacc aggacaaggc tgcccccgag
     1801 atggtttctc tgctgaagag gaataactgt gggaaagccc tggacatcgc ccgccaggcc
     1861 cgagacatgc tgggggggaa tgggatttct gacgagtatc acgtgatccg gcacgccatg
     1921 aacctggagg ccgtgaacac ctacgaaggt acacatgaca ttcacgccct gatccttggg
     1981 agagctatca cgggaatcca ggcgttcacg gccagcaagc tcgagtaccc atacgacgta
     2041 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     2101 ccagactacg catagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt
     2161 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2221 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2281 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2341 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2401 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2461 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2521 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     2581 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2641 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2701 tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga
     2761 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     2821 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     2881 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     2941 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3001 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3181 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     3421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3841 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3901 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3961 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4021 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4081 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4141 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4201 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4261 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4321 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4381 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4441 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4501 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4561 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4621 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4681 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4741 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4801 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4861 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4921 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4981 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5101 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5581 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5821 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6241 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7141 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7201 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7261 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7321 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
