LOCUS       V025766                 7624 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025766
VERSION     V025766
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             907..2430
                     /label="ALDH3A2(NM_001031806)"
                     /note="ALDH3A2(NM_001031806)"
                     /gene="ALDH3A2"
     CDS             2467..3186
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    3224..3448
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3494..3922
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3936..4265
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4332..5126
                     /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    5300..5421
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5470..5486)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5494..5510
                     /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(5518..5548)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5563..5584
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5872..6457)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6628..7488)
                     /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(7489..7593)
                     /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 gccaccatgg agctcgaagt ccggcgggtc cgacaggcgt tcctgtccgg ccggtcgcga
      961 cctctgcggt ttcggctgca gcagctggag gccctgcgga ggatggtgca ggagcgcgag
     1021 aaggatatcc tgacggccat cgccgccgac ctgtgcaaga gtgaattcaa tgtgtacagt
     1081 caggaagtca ttactgtcct tggggaaatt gattttatgc ttgagaatct tcctgaatgg
     1141 gttactgcta aaccagttaa gaagaacgtg ctcaccatgc tggatgaggc ctatattcag
     1201 ccacagcctc tgggagtggt gctgataatc ggagcttgga attacccctt cgttctcacc
     1261 attcagccac tgataggagc catcgctgca ggaaatgctg tgattataaa gccttctgaa
     1321 ctgagtgaaa atacagccaa gatcttggca aagcttctcc ctcagtattt agaccaggat
     1381 ctctatattg ttattaatgg tggtgttgag gaaaccacgg agctcctgaa gcagcgattt
     1441 gaccacattt tctatacggg aaacactgcg gttggcaaaa ttgtcatgga agctgctgcc
     1501 aagcatctga cccctgtgac tcttgaactg ggagggaaaa gtccatgtta tattgataaa
     1561 gattgtgacc tggacattgt ttgcagacgc ataacctggg gaaaatacat gaattgtggc
     1621 caaacctgca ttgcacccga ctatattctc tgtgaagcat ccctccaaaa tcaaattgta
     1681 tggaagatta aggaaacagt gaaggaattt tatggagaaa atataaaaga gtctcctgat
     1741 tatgaaagga tcatcaatct tcgtcatttt aagaggatac taagtttgct tgaaggacaa
     1801 aagatagctt ttggtgggga gactgatgag gccacacgct acatagcccc aacagtactt
     1861 accgatgttg atcctaaaac caaggtgatg caagaagaaa tttttggacc aattcttcca
     1921 atagtgcctg tgaaaaatgt agatgaggcc ataaatttca taaatgaacg tgaaaagcct
     1981 ctggctcttt atgtattttc gcataaccat aagctcatca aacggatgat tgatgagaca
     2041 tccagtggag gtgtcacagg caatgacgtc attatgcact tcacgctcaa ctctttccca
     2101 tttggaggag tgggttccag tgggatggga gcttatcacg gaaaacatag ttttgatact
     2161 ttttctcatc agcgtccctg tttattaaaa agtttaaaga gagaaggtgc taacaaactc
     2221 agatatcctc ccaacagcca gtcaaaggtg gattggggaa aattttttct cttgaaacgg
     2281 ttcaacaaag aaaaactcgg tctcctgttg ctcactttcc tgggtattgt agccgctgtg
     2341 cttgtcaaga aataccaagc tgtgctgagg agaaaggccc tgttgatttt tctggtagtt
     2401 cacagactgc gttggtccag taagcagaga ggatccggcg gtggaggaag cggcggaggt
     2461 ggatctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2521 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2581 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2641 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2701 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2761 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2821 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2881 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2941 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3001 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3061 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3121 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3181 aagtagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3241 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3301 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3361 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3421 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3481 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3541 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3601 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3661 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3721 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3781 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3841 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3901 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4141 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4201 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4261 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4321 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4381 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4441 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4501 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4561 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4621 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4681 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4741 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4801 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4861 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4921 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4981 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5041 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5101 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5161 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5221 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5281 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5341 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5401 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5461 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6541 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6601 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6661 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6721 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6781 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6841 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6901 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6961 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7021 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7081 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7141 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7201 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7261 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7321 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7381 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7441 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7501 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7561 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7621 cgtc
//
