LOCUS       V023862                 8703 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023862
VERSION     V023862
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        236..615
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        616..819
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        864..882
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             965..3490
                     /label="UNC5A(NM_133369)"
                     /note="UNC5A(NM_133369)"
                     /gene="UNC5A"
     CDS             3533..4252
                     /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    4304..4528
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4574..5002
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5016..5345
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5412..6206
                     /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    6380..6501
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6550..6566)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6574..6590
                     /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(6598..6628)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6643..6664
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6952..7537)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7708..8568)
                     /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(8569..8673)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc
      961 caccatggcc gtccggcccg gcctgtggcc agcgctcctg ggcatagtcc tcgccgcttg
     1021 gctccgcggc tcgggtgccc agcagagtgc caccgtggcc aacccagtgc ctggtgccaa
     1081 cccggacctg cttccccact tcctggtgga gcccgaggat gtgtacatcg tcaagaacaa
     1141 gccagtgctg cttgtgtgca aggccgtgcc cgccacgcag atcttcttca agtgcaacgg
     1201 ggagtgggtg cgccaggtgg accacgtgat cgagcgcagc acagacggga gcagtgggct
     1261 gcccaccatg gaggtccgca ttaatgtctc aaggcagcag gtcgagaagg tgttcgggct
     1321 ggaggaatac tggtgccagt gcgtggcatg gagctcctcg ggcaccacca agagtcagaa
     1381 ggcctacatc cgcatagcct atttgcgcaa gaacttcgag caggagccgc tggccaagga
     1441 ggtgtccctg gagcagggca tcgtgctgcc ctgccgtcca ccggagggca tccctccagc
     1501 cgaggtggag tggctccgga acgaggacct ggtggacccg tccctggacc ccaatgtata
     1561 catcacgcgg gagcacagcc tggtggtgcg acaggcccgc cttgctgaca cggccaacta
     1621 cacctgcgtg gccaagaaca tcgtggcacg tcgccgcagc gcctccgctg ctgtcatcgt
     1681 ctacgtggac ggcagctgga gcccgtggag caagtggtcg gcctgtgggc tggactgcac
     1741 ccactggcgg agccgtgagt gctctgaccc agcaccccgc aacggagggg aggagtgcca
     1801 gggcactgac ctggacaccc gcaactgtac cagtgacctc tgtgtacaca ctgcttctgg
     1861 ccctgaggac gtggccctct atgtgggcct catcgccgtg gccgtctgcc tggtcctgct
     1921 gctgcttgtc ctcatcctcg tttattgccg gaagaaggag gggctggact cagatgtggc
     1981 tgactcgtcc attctcacct caggcttcca gcccgtcagc atcaagccca gcaaagcaga
     2041 caacccccat ctgctcacca tccagccgga cctcagcacc accaccacca cctaccaggg
     2101 cagtctctgt ccccggcagg atgggcccag ccccaagttc cagctcacca atgggcacct
     2161 gctcagcccc ctgggtggcg gccgccacac actgcaccac agctctccca cctctgaggc
     2221 cgaggagttc gtctcccgcc tctccaccca gaactacttc cgctccctgc cccgaggcac
     2281 cagcaacatg acctatggga ccttcaactt cctcgggggc cggctgatga tccctaatac
     2341 aggaatcagc ctcctcatcc ccccagatgc cataccccga gggaagatct atgagatcta
     2401 cctcacgctg cacaagccgg aagacgtgag gttgccccta gctggctgtc agaccctgct
     2461 gagtcccatc gttagctgtg gaccccctgg cgtcctgctc acccggccag tcatcctggc
     2521 tatggaccac tgtggggagc ccagccctga cagctggagc ctgcgcctca aaaagcagtc
     2581 gtgcgagggc agctgggagg atgtgctgca cctgggcgag gaggcgccct cccacctcta
     2641 ctactgccag ctggaggcca gtgcctgcta cgtcttcacc gagcagctgg gccgctttgc
     2701 cctggtggga gaggccctca gcgtggctgc cgccaagcgc ctcaagctgc ttctgtttgc
     2761 gccggtggcc tgcacctccc tcgagtacaa catccgggtc tactgcctgc atgacaccca
     2821 cgatgcactc aaggaggtgg tgcagctgga gaagcagctg gggggacagc tgatccagga
     2881 gccacgggtc ctgcacttca aggacagtta ccacaacctg cgcctatcca tccacgatgt
     2941 gcccagctcc ctgtggaaga gtaagctcct tgtcagctac caggagatcc ccttttatca
     3001 catctggaat ggcacgcagc ggtacttgca ctgcaccttc accctggagc gtgtcagccc
     3061 cagcactagt gacctggcct gcaagctgtg ggtgtggcag gtggagggcg acgggcagag
     3121 cttcagcatc aacttcaaca tcaccaagga cacaaggttt gctgagctgc tggctctgga
     3181 gagtgaagcg ggggtcccag ccctggtggg ccccagtgcc ttcaagatcc ccttcctcat
     3241 tcggcagaag ataatttcca gcctggaccc accctgtagg cggggtgccg actggcggac
     3301 tctggcccag aaactccacc tggacagcca tctcagcttc tttgcctcca agcccagccc
     3361 cacagccatg atcctcaacc tgtgggaggc gcggcacttc cccaacggca acctcagcca
     3421 gctggctgca gcagtggctg gactgggcca gccagacgct ggcctcttca cagtgtcgga
     3481 ggctgagtgc ggaggagggg gaagcggagg cggagggaga ccggtcgcca ccatggtgag
     3541 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3601 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3661 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3721 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3781 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3841 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3901 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3961 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4021 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4081 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4141 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4201 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     4261 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     4321 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4381 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4441 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4501 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4561 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4621 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4681 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4741 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4801 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4861 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4921 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4981 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5101 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5161 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5221 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5281 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5341 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5401 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5461 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5521 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5581 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5641 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5701 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5761 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5821 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5881 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5941 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6001 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6061 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6121 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6181 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6241 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6301 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6361 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6421 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6481 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     7501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7741 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7801 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7861 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7921 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7981 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8041 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8101 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8161 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8221 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8281 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8341 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8401 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8461 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8521 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8581 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8641 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8701 cgt
//
