LOCUS       V047290                 8781 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047290
VERSION     V047290
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..3461
                     /label="Aldh1l1(NM_027406)"
                     /note="Aldh1l1(NM_027406)"
                     /gene="Aldh1l1"
     CDS             3468..3533
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3557..4145
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4211..4259
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4461..4889
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4903..5232
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5299..6093
                     /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    6269..6390
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6439..6455)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6463..6479
                     /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(6487..6517)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6532..6553
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6841..7429)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7600..8460)
                     /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(8461..8565)
                     /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 ccaccatgaa gattgcagta atcgggcaga
      781 gcctgtttgg ccaggaggtt tactgccagc taaggaagga gggccatgag gtggtgggtg
      841 tgttcaccat cccagacaag gatgggaaag cagatcccct gggcctggag gccgagaagg
      901 atggcgtgcc cgtgtttaag ttccctcggt ggcgggctcg aggacaggct ctgcccgagg
      961 tcgtggccaa gtaccaggct ctgggtgcag aactcaacgt gctccccttc tgcagccagt
     1021 tcatacccat ggaggtcatc aatgcccccc ggcatggctc catcatctac catccatctc
     1081 tgctgcccag gcaccgaggg gcctcagcca tcaactggac cctcattcat ggagataaga
     1141 aaggaggctt cactatcttc tgggctgatg acggtctgga cactggtgac cttctgctgc
     1201 agaaggagtg tgacgtgctt ccagatgaca ccgtgagcac gctgtacaac cgtttcctct
     1261 tcccagaggg catcaaaggg atggttcagg ctgtgcgact gattgcagag ggcacagccc
     1321 cacgccgtcc ccagcctgag gaaggagcca cctatgaggg cattcagaaa aaggagacag
     1381 ccatgatcaa ctgggaccag ccagcagagg ccattcacaa ctggatccgt gggaatgaca
     1441 aggtgccagg tgcctggaca gaggcttgtg ggcagaagct gacgtttttc aactcgacac
     1501 tcaacacttc aggactggtg gctcagggag aagctctacc catcccagga gcccatcgtc
     1561 caggcctagt caccaaagcg ggactcatcc tctttgggaa tgatgataga atgttgctgg
     1621 tgaagaatat ccagctggaa gatggcaaga tgatgccagc ctcccagttc ttcaaggggt
     1681 ctgctagcag tgccctggag ctgaccgagg aggagctggc cacagcagag gccgtgcgga
     1741 gctcttggat gcgaattctg cccaatgtcc cagaggtaga agactctaca gatttcttca
     1801 agtcaggagc tgcatccgta gatgttgtga ggctggtgga ggaggtgaag gagctgtgtg
     1861 acgggctgga gttagaaaat gaggatgttt acatggccac caccttcggg gacttcatcc
     1921 agctcctagt gaggaagttg agaggggagg acggcgagag cgagtgtgtc attaactacg
     1981 tggaaaaggc agtgaagaaa ctgactctcc aaatgcccta ccagctcttc ataggcggcg
     2041 agtttgtgga tgctgagggc gcgaagacct acagtaccat aaacccaacg gatggaagtg
     2101 tcatctgcca ggtgtctcta gctcaggtga gtgatgttga caaggcggtg gcagcagcga
     2161 aggaggcctt tgagaatgga ctgtggggaa agataaatgc gcgtgaccgg ggccggctcc
     2221 tgtacaggtt ggcggacctc atggagcagc accaggaaga gctagccacc attgaggccc
     2281 tggacgcagg tgccgtctac acgctggccc tgaagacgca tgtgggcatg tccatccaga
     2341 ccttccgata cttcgctggc tggtgtgata agatccaggg tgccaccatc cccatcaacc
     2401 aggctagacc caaccgcaac ctgaccttga ccaagaagga acctgttggg gtctgtggta
     2461 ttgtcatccc ctggaactat cccttaatga tgctgtcctg gaagacagca gcctgcctgg
     2521 ctgccgggaa caccgtggtg atcaagcctg cccaggtgac cccactcaca gccttgaagt
     2581 ttgcagagct gacactgaag gctggcattc ccaagggtgt ggtcaacatc ctcccaggat
     2641 ctggctcgct ggttggccag agactctcag accaccctga tgtgaggaaa atagggttca
     2701 caggctccac ggaggtggga aaacacatca tgaaaagctg tgccctgagt aatgtgaaga
     2761 aggtctccct ggagctgggt ggaaagtcac cccttatcat ctttgctgac tgtgacctca
     2821 acaaagctgt gcagatgggc atgagctccg ttttcttcaa caaaggggag aactgcattg
     2881 cggcaggccg gctctttgtg gaggactcca tccatgacca gtttgtgcag aaagtggtgg
     2941 aggaagtagg gaagatgaaa atcggcaacc ccctggacag agataccaac catggcccgc
     3001 agaaccatga ggcccacctg aggaagctgg tggagtattg ccaacgtggt gtgaaggaag
     3061 gggccacact ggtctgtggt gggaaccaag tcccaaggcc aggcttcttc tttcagccaa
     3121 ccgttttcac agacgtggag gaccacatgt acatcgctaa ggaggagtcc ttcgggccca
     3181 tcatgatcat ctctcggttt gctgatgggg acgtggatgc agtgctatct cgggccaatg
     3241 ctacagaatt tggcctggcc tctggtgtct tcactcggga tatcaacaag gccctgtatg
     3301 tcagtgacaa actgcaggcg ggcactgtgt ttgtcaacac atacaacaag accgatgtgg
     3361 ccgcaccttt tggaggattc aagcagtctg gatttggcaa agacctggga gaggcggccc
     3421 tgaatgagta cctgcggatc aagactgtga cttttgagta cggatccgac tacaaagacc
     3481 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     3541 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3601 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3661 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3721 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3781 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3841 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3901 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3961 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4021 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4081 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4141 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4201 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4261 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4321 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4381 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     4441 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4501 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4561 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     4621 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4681 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4741 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4801 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     4861 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5281 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5341 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5401 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5461 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5521 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5581 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5641 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5701 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5761 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5821 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5881 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5941 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6001 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6061 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6121 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6181 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6241 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6421 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7021 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7261 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8581 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     8641 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     8701 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     8761 tgcatgaaga atctgcttag g
//
