LOCUS       V035228                 7632 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035228
VERSION     V035228
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..2312
                     /label="Aldh2(NM_009656)"
                     /note="Aldh2(NM_009656)"
                     /gene="Aldh2"
     CDS             2319..2384
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2408..2996
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3062..3110
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3312..3740
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3754..4083
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4150..4944
                     /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    5120..5241
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5290..5306)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5314..5330
                     /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(5338..5368)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5383..5404
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5692..6280)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6451..7311)
                     /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(7312..7416)
                     /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 ccaccatgct gcgcgccgca ctcaccactg
      781 tccgccgcgg accgcgcctg agccgcctgt tgtccgccgc cgccaccagc gcggtgccag
      841 cccccaacca tcagcctgag gtcttctgca accagatctt cattaacaat gagtggcacg
      901 acgccgtcag caggaaaaca tttcccaccg tcaacccttc cacaggggag gtcatctgcc
      961 aggtggccga agggaacaag gaggacgtag acaaggcagt gaaggctgct cgtgcagcct
     1021 tccagctggg ctcgccctgg cgccgcatgg atgcatctga ccggggccgg ctgttgtacc
     1081 gattggcgga tctcattgaa cgggaccgga cctacctagc ggccttggag accctggaca
     1141 acggcaagcc ttatgtcatc tcgtacctgg tggatttgga catggtcctg aaatgtctcc
     1201 gctattacgc tggctgggct gacaagtacc atgggaaaac cattcccatc gacggcgact
     1261 tcttcagcta tacccgccat gagcctgtgg gcgtgtgtgg acagatcatt ccgtggaact
     1321 tcccgctcct gatgcaagca tggaaactgg gcccagccct ggcaaccggg aacgtggtgg
     1381 tgatgaaggt ggccgagcag acaccgctca ccgcgctcta cgtggccaac ttgatcaagg
     1441 aggcaggctt tccccctggc gtggtcaata tcgttcccgg attcggccct accgccgggg
     1501 ctgccatcgc atcccatgag ggtgtggaca aagtggcgtt cacaggctcc acggaggttg
     1561 gtcacctaat ccaggtggcc gccgggagca gcaacctcaa gagagtaacc ctggagctgg
     1621 ggggaaagag tcccaacatc atcatgtccg acgctgacat ggactgggct gtggagcagg
     1681 cccactttgc cctgttcttc aaccagggcc agtgctgctg cgcaggctcc cggaccttcg
     1741 tgcaggagaa tgtgtatgac gaattcgtgg aacgcagcgt ggctcgggcc aagtctcggg
     1801 tggtggggaa ccccttcgac agccggacgg agcaggggcc tcaggtggat gaaactcagt
     1861 ttaagaagat cctcggctac atcaaatcgg gacaacaaga aggggcgaag ctgctgtgtg
     1921 gtgggggcgc tgccgcggac cgtggctact ttatccagcc caccgtgttc ggggacgtaa
     1981 aagacggcat gaccattgcc aaggaggaga tctttggacc agtgatgcaa atcctcaaat
     2041 tcaagaccat cgaggaggtt gtggggcggg ccaatgattc taagtatggg ctggcagccg
     2101 ccgtcttcac aaaggacctg gataaagcca attacctgtc ccaagctctg caggctggca
     2161 ctgtgtggat caactgctac gatgtgtttg gggcccagtc tccatttggg ggctataaga
     2221 tgtcagggag tggcagggag ctgggcgagt atggcctgca ggcgtacaca gaagtgaaga
     2281 cggttactgt caaagtgcca cagaagaact cgggatccga ctacaaagac catgacggtg
     2341 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2401 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2461 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2521 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2581 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2641 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2701 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2761 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2821 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2881 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2941 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3001 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3061 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3121 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3181 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3241 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3301 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3361 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3421 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3481 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3541 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3601 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3661 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3721 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3841 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3901 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3961 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4021 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4081 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4141 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4201 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4261 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4321 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4381 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4441 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4501 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4561 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4621 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4681 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4741 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4801 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4861 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4921 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     4981 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5041 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5101 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5161 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5221 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5281 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5341 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5401 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5461 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5521 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5881 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6121 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6541 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6601 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6661 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6721 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6781 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6841 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6901 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6961 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7021 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7081 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7141 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7201 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7261 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7441 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7501 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7561 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7621 aatctgctta gg
//
