LOCUS       V041124                 8502 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041124
VERSION     V041124
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             887..3271
                     /label="ALDH18A1(NM_002860)"
                     /note="ALDH18A1(NM_002860)"
                     /gene="ALDH18A1"
     CDS             3308..4018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    4069..4293
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4339..4767
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4781..5110
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5177..5971
                     /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    6145..6266
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6315..6331)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6339..6355
                     /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(6363..6393)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6408..6429
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6717..7302)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7473..8333)
                     /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(8334..8438)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcaac ttaaggatcc gccaccatgt tgagtcaagt
      901 ttaccgctgt gggttccagc ccttcaacca acatcttctg ccctgggtca agtgtacaac
      961 cgtcttcaga tctcattgta tccagccttc agtcatcaga catgttcgtt cttggagcaa
     1021 catcccgttt atcactgtac ccctcagtcg tacacatggc aagtccttcg cccaccgcag
     1081 tgagctgaag catgccaaga gaatcgtggt gaagctcggc agtgccgtgg tgacccgagg
     1141 ggatgaatgt ggcctggccc tggggcgctt ggcatctatt gttgagcagg tatcagtgct
     1201 gcagaatcag ggcagagaga tgatgctggt gaccagtgga gccgtagcct ttggcaaaca
     1261 acgcttgcgc catgagatcc ttctgtctca gagcgtgcgg caggccctcc actcggggca
     1321 gaaccagctg aaagaaatgg caattccagt cttagaggca cgagcctgtg cagctgccgg
     1381 acagagtggg ctgatggcct tgtatgaggc tatgtttacc cagtacagca tctgtgctgc
     1441 ccagattttg gtgaccaatt tggatttcca tgatgagcag aagcgccgga acctcaatgg
     1501 aacacttcat gaactcctta gaatgaacat tgtccccatt gtcaacacaa atgatgctgt
     1561 tgtcccccca gctgagccca acagtgacct gcagggggta aatgttatta gtgttaaaga
     1621 taatgatagc ctggctgccc gactggctgt ggaaatgaaa actgatctct tgattgttct
     1681 ttcagatgta gaaggccttt ttgacagccc cccaggttca gatgatgcaa agcttattga
     1741 tatattttat cccggagatc agcagtctgt gacatttgga accaagtcta gagtgggaat
     1801 gggtggcatg gaagccaagg tgaaagcagc cctctgggct ttgcaaggtg gcacttctgt
     1861 tgttattgcc aatggaaccc acccaaaggt gtctgggcac gtcatcacag acattgtgga
     1921 ggggaagaaa gttggtacct tcttttcaga agtaaagcct gcaggcccta ctgttgagca
     1981 gcagggagaa atggcgcgat ctggaggaag gatgttggcc accttggaac ctgagcagag
     2041 agcagaaatt atccatcatc tggctgatct gttgacggac cagcgtgatg agatcctgtt
     2101 agccaacaaa aaagacttgg aggaggcaga ggggagactt gcagctcctc tgctgaaacg
     2161 tttaagcctc tccacatcca aattgaacag cctggccatc ggtctgcgac agatcgcagc
     2221 ctcctcccag gacagcgtgg gacgtgtttt gcgccgcacc cgaatcgcca aaaacttgga
     2281 actggaacaa gtgactgtcc caattggagt tctgctggtg atctttgaat ctcgtcctga
     2341 ctgtctaccc caggtggcag ctttggctat cgcaagtggc aatggcttgt tactcaaagg
     2401 agggaaggag gctgcacaca gcaaccggat tctccacctc ctgacccagg aggctctctc
     2461 aatccatgga gtcaaggagg ccgtgcaact ggtgaatacc agagaagaag ttgaagatct
     2521 ttgccgccta gacaaaatga tagatctgat cattccacgt ggctcttccc agctggtcag
     2581 agacatccag aaagctgcta aggggattcc agtgatgggg cacagcgaag ggatctgtca
     2641 catgtatgtg gattccgagg ccagtgttga taaggtcacc aggctagtca gagactctaa
     2701 atgtgaatat ccagctgcct gtaatgcttt ggagactttg ttaatccacc gggatctgct
     2761 caggacacca ttatttgacc agatcattga tatgctgaga gtggaacagg taaaaattca
     2821 tgcaggcccc aaatttgcct cctatctgac cttcagcccc tccgaagtga agtcactccg
     2881 aactgagtat ggggacctgg aattatgcat tgaagtagtg gacaacgttc aggatgccat
     2941 tgaccacatc cacaagtatg gcagctccca cacggatgtc atcgtcacag aggacgaaaa
     3001 cacagcggag ttcttcctgc agcacgtaga cagtgcctgt gtgttctgga atgccagcac
     3061 tcgcttttct gatggttacc gctttggact gggagctgaa gtgggaatca gtacatcgag
     3121 aatccacgcc cggggaccag taggacttga gggactgctt actactaagt ggctgctgcg
     3181 agggaaggac cacgtggtct cagatttctc agagcatgga agtttaaaat atcttcatga
     3241 gaacctccct attcctcaga gaaacaccaa cgaattcggt ggaggcggat caggaggcgg
     3301 tggatctatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     3361 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     3421 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     3481 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     3541 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     3601 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     3661 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     3721 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     3781 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     3841 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     3901 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3961 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaggc
     4021 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     4081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5161 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5221 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5281 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5341 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5401 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5461 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5521 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5581 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5641 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5701 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5761 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5821 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5881 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5941 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6001 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6061 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6121 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6241 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8461 cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
