LOCUS       V041217                 7626 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041217
VERSION     V041217
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2306
                     /label="ALDH2(NM_000690)"
                     /note="ALDH2(NM_000690)"
                     /gene="ALDH2"
     CDS             2313..2378
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2402..2990
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3056..3104
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3306..3734
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3748..4077
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4144..4938
                     /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    5114..5235
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5284..5300)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5308..5324
                     /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(5332..5362)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5377..5398
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5686..6274)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6445..7305)
                     /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(7306..7410)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtt gcgcgctgcc gcccgcttcg
      781 ggccccgcct gggccgccgc ctcttgtcag ccgccgccac ccaggccgtg cctgccccca
      841 accagcagcc cgaggtcttc tgcaaccaga ttttcataaa caatgaatgg cacgatgccg
      901 tcagcaggaa aacattcccc accgtcaatc cgtccactgg agaggtcatc tgtcaggtag
      961 ctgaagggga caaggaagat gtggacaagg cagtgaaggc cgcccgggcc gccttccagc
     1021 tgggctcacc ttggcgccgc atggacgcat cacacagggg ccggctgctg aaccgcctgg
     1081 ccgatctgat cgagcgggac cggacctacc tggcggcctt ggagaccctg gacaatggca
     1141 agccctatgt catctcctac ctggtggatt tggacatggt cctcaaatgt ctccggtatt
     1201 atgccggctg ggctgataag taccacggga aaaccatccc cattgacgga gacttcttca
     1261 gctacacacg ccatgaacct gtgggggtgt gcgggcagat cattccgtgg aatttcccgc
     1321 tcctgatgca agcatggaag ctgggcccag ccttggcaac tggaaacgtg gttgtgatga
     1381 aggtagctga gcagacaccc ctcaccgccc tctatgtggc caacctgatc aaggaggctg
     1441 gctttccccc tggtgtggtc aacattgtgc ctggatttgg ccccacggct ggggccgcca
     1501 ttgcctccca tgaggatgtg gacaaagtgg cattcacagg ctccactgag attggccgcg
     1561 taatccaggt tgctgctggg agcagcaacc tcaagagagt gaccttggag ctggggggga
     1621 agagccccaa catcatcatg tcagatgccg atatggattg ggccgtggaa caggcccact
     1681 tcgccctgtt cttcaaccag ggccagtgct gctgtgccgg ctcccggacc ttcgtgcagg
     1741 aggacatcta tgatgagttt gtggagcgga gcgttgcccg ggccaagtct cgggtggtcg
     1801 ggaacccctt tgatagcaag accgagcagg ggccgcaggt ggatgaaact cagtttaaga
     1861 agatcctcgg ctacatcaac acggggaagc aagagggggc gaagctgctg tgtggtgggg
     1921 gcattgctgc tgaccgtggt tacttcatcc agcccactgt gtttggagat gtgcaggatg
     1981 gcatgaccat cgccaaggag gagatcttcg ggccagtgat gcagatcctg aagttcaaga
     2041 ccatagagga ggttgttggg agagccaaca attccacgta cgggctggcc gcagctgtct
     2101 tcacaaagga tttggacaag gccaattacc tgtcccaggc cctccaggcg ggcactgtgt
     2161 gggtcaactg ctatgatgtg tttggagccc agtcaccctt tggtggctac aagatgtcgg
     2221 ggagtggccg ggagttgggc gagtacgggc tgcaggcata cactgaagtg aaaactgtca
     2281 cagtcaaagt gcctcagaag aactcaggat ccgactacaa agaccatgac ggtgattata
     2341 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2401 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2461 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2521 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2581 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2641 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2701 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2761 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2821 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2881 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2941 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3001 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3061 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3121 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3181 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3241 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3301 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3601 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3661 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3721 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3961 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4021 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4081 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4141 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4201 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4261 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4321 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4381 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4441 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4501 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4561 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4621 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4681 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4741 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4801 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4861 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4921 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4981 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5041 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5101 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5161 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5221 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5281 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5341 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5401 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5461 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5521 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5581 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5641 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5701 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5761 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5821 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5881 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5941 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6001 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6061 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6121 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6181 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6241 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6301 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6361 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6421 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6481 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6541 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6601 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6661 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6721 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6781 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6841 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6901 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6961 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7021 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7081 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7141 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7201 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7261 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7381 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7441 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7501 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7561 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7621 cttagg
//
