LOCUS       V047172                 7879 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047172
VERSION     V047172
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             925..3630
                     /label="ALDH1L1(NM_012190)"
                     /note="ALDH1L1(NM_012190)"
                     /gene="ALDH1L1"
     CDS             3643..3708
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3743..3967
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4013..4441
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4455..4784
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4796..5395
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5555..5676
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5725..5741)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5749..5765
                     /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(5773..5803)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5818..5839
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6127..6712)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6883..7743)
                     /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(7744..7848)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcgc caccatgaag attgcagtga ttggacagag cctgtttggc
      961 caggaagttt actgccacct gaggaaggag ggccacgaag tggtgggtgt gttcactgtt
     1021 ccagacaagg atggaaaggc cgaccccctg ggtctggaag ctgagaagga tggagtgccg
     1081 gtattcaagt actcccggtg gcgtgcaaaa ggacaggctt tgcctgatgt ggtggcaaaa
     1141 taccaggctt tgggggccga gctcaacgtc ctgcccttct gcagccaatt catccccatg
     1201 gagataatca gtgccccccg gcatggctcc atcatctatc acccgtcact gctccctagg
     1261 caccgagggg cctcggccat caactggacc ctcattcacg gagataagaa aggggggttt
     1321 tccatcttct gggcggatga tggtctggac accggagacc tgctgctgca gaaggagtgt
     1381 gaggtgctcc cggacgacac cgtgagcacg ctgtacaacc gcttcctctt ccctgaaggc
     1441 atcaaaggga tggtgcaggc cgtgaggctg atcgctgagg gcaaagcccc cagactccct
     1501 cagcctgagg aaggagccac ctatgagggg attcagaaga aggagacagc caagatcaac
     1561 tgggaccagc cggcagaggc cattcacaac tggatccgcg ggaacgacaa ggtgccggga
     1621 gcctggacag aggcctgtga acagaaactg acatttttca actcaacgct gaacacttca
     1681 ggcctggtgc ccgagggaga cgctttgccc atcccaggag cccatcggcc aggggtggtc
     1741 accaaagcag gactcatcct ctttgggaat gatgacaaaa tgctgctggt gaagaatatt
     1801 cagctggagg atggcaaaat gatcctggcc tcgaacttct ttaagggggc agccagcagt
     1861 gtccttgagc tgacagaggc agagctggtt actgcggagg ctgtgcggag tgtttggcag
     1921 cggatcctcc ccaaagtcct ggaggttgaa gactccactg atttcttcaa gtcaggggcc
     1981 gcgtctgtgg acgttgtgag gctggtggag gaagtgaagg agctgtgtga tggcctggag
     2041 ttagaaaatg aagatgtgta catggcatcc acctttgggg acttcatcca gctgttagtg
     2101 aggaagctgc gaggggacga tgaggagggc gagtgcagca ttgactacgt ggaaatggca
     2161 gtgaacaagc gcactgtccg catgccccac cagctcttca ttggggggga gttcgtggat
     2221 gccgagggcg ccaagacctc tgagaccatc aatcccaccg atggaagtgt catctgccag
     2281 gtatccctgg cccaagtcac cgacgtcgac aaggcagtgg ccgcagccaa ggatgccttt
     2341 gagaatggac ggtgggggaa gatcagtgcg cgggaccggg gccggctgat gtacaggttg
     2401 gcagatctca tggagcagca ccaggaggag ctggccacca ttgaggccct ggatgcgggt
     2461 gccgtctaca cgctggccct gaagacccac gtgggcatgt ccatccagac cttccgctac
     2521 tttgctggct ggtgtgacaa gatccagggc tccaccatcc ccatcaacca ggccagaccc
     2581 aaccgcaacc tgaccttgac caggaaggag cctgttgggg tttgtggcat catcatcccc
     2641 tggaactatc ccctgatgat gctgtcctgg aagacagctg cctgcctggc tgccgggaac
     2701 acagtggtga tcaagcctgc tcaggtgacc ccactcacag ccttgaagtt tgcagagctg
     2761 acattaaagg ccggcattcc caaaggtgtg gttaacgtcc tcccaggatc tggctccctg
     2821 gtcggccaga gactctcaga ccatcctgat gtgaggaaaa tcgggttcac aggctccaca
     2881 gaggtgggca agcacatcat gaaaagctgt gccataagta acgtgaagaa ggtgtccctg
     2941 gaactgggcg ggaagtcacc cctcatcatc tttgctgact gtgacctcaa caaggctgtg
     3001 cagatgggga tgagttctgt tttcttcaac aaaggagaga attgcattgc agcaggccga
     3061 ctctttgtgg aggactccat tcatgatgag ttcgtgcgga gagtggtaga agaggtgcgg
     3121 aagatgaagg tgggcaaccc gctggacagg gacaccgacc acgggccgca gaatcaccat
     3181 gcccaccttg tgaagctgat ggagtactgc cagcatggcg tgaaggaagg ggccacactg
     3241 gtctgcggcg ggaatcaggt ccctcggcca gggttcttct ttgagccaac tgttttcaca
     3301 gacgtggaag accacatgtt catagccaag gaggagtcct tcgggcctgt catgatcatc
     3361 tctcggtttg ctgatgggga cttggatgcc gtgctgtctc gggccaatgc cacggaattt
     3421 ggcctggctt ctggtgtctt caccagggac atcaacaagg ccctgtatgt cagtgacaag
     3481 ctccaggcag gcactgtgtt tgtcaacacg tacaacaaga ccgacgtggc cgctcccttc
     3541 ggaggattca aacagtctgg atttggcaaa gatctaggag aggcggctct gaacgagtac
     3601 ctgcgggtca agacagtgac cttcgaatac ctcgagaccg gtgactacaa agaccatgac
     3661 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagtg accggtaagt
     3721 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3781 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3841 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3901 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3961 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4021 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4081 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4141 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4201 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4261 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4321 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4381 ttggccattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4441 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4501 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4621 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4681 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4741 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggatgac
     4801 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4861 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4921 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4981 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5041 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5101 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5161 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5221 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5281 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5341 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga
     5401 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5461 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5521 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5581 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5641 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5701 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5761 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5821 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5881 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5941 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6001 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6061 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6661 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7861 aaaagtgcca cctgacgtc
//
