LOCUS       V027798                 6979 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027798
VERSION     V027798
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..1635
                     /label="SIRT5(NM_012241)"
                     /note="SIRT5(NM_012241)"
                     /gene="SIRT5"
     CDS             1642..1731
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1755..2343
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2409..2457
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2659..3087
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3101..3430
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3497..4291
                     /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    4467..4588
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4637..4653)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4661..4677
                     /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(4685..4715)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4730..4751
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5039..5627)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5798..6658)
                     /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(6659..6763)
                     /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 ccaccatgcg acctctccag
      721 attgtcccaa gtcgattgat ttcccagcta tattgtggcc tgaagcctcc agcgtccaca
      781 cgaaaccaga tttgcctgaa aatggctcgg ccaagttcaa gtatggcaga ttttcgaaag
      841 ttttttgcaa aagcaaagca catagtcatc atctcaggag ctggtgttag tgcagaaagt
      901 ggtgttccga ccttcagagg agctggaggt tattggagaa aatggcaagc ccaggacctg
      961 gcgactcccc tggcctttgc ccacaacccg tcccgggtgt gggagttcta ccactaccgg
     1021 cgggaggtca tggggagcaa ggagcccaac gccgggcacc gcgccatagc cgagtgtgag
     1081 acccggctgg gcaagcaggg ccggcgagtc gtggtcatca cccagaacat cgatgagctg
     1141 caccgcaagg ctggcaccaa gaaccttctg gagatccatg gtagcttatt taaaactcga
     1201 tgtacctctt gtggagttgt ggctgagaat tacaagagtc caatttgtcc agctttatca
     1261 ggaaaaggtg ctccagaacc tggaactcaa gatgccagca tcccagttga gaaacttccc
     1321 cggtgtgaag aggcaggctg cgggggcttg ctgcgacctc acgtcgtgtg gtttggagaa
     1381 aacctggatc ctgccattct ggaggaggtt gacagagagc tcgcccactg tgatttatgt
     1441 ctagtggtgg gcacttcctc tgtggtgtac ccagcagcca tgtttgcccc ccaggtggct
     1501 gccaggggcg tgccagtggc tgaatttaac acggagacca ccccagctac gaacagattc
     1561 aggtttcatt tccagggacc ctgtggaacg actcttcctg aagcccttgc ctgtcatgaa
     1621 aatgaaactg tttctctcga ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa
     1681 ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt atagtgaggg
     1741 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2161 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2341 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     2401 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     2461 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     2521 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     2581 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     2641 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2701 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2761 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     2821 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2881 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2941 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3001 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     3061 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3181 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3241 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3301 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3361 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3421 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3481 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3541 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3601 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3661 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3721 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3781 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3841 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3901 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3961 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4021 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4081 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4141 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4201 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4261 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     4321 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4381 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4441 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4561 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4621 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4681 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4741 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4801 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4861 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4921 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4981 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5041 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5101 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5161 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5221 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5281 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5341 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5401 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5461 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5521 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5581 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5641 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5701 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5761 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5821 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5881 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5941 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6001 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6061 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6121 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6181 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6241 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6301 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6361 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6421 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6481 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6541 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6601 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6781 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac
     6841 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6901 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6961 catgaagaat ctgcttagg
//
