LOCUS       V027907                 7837 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027907
VERSION     V027907
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             874..1590
                     /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"
     CDS             1642..3327
                     /label="SPTLC2(NM_004863)"
                     /note="SPTLC2(NM_004863)"
                     /gene="SPTLC2"
     polyA_signal    3404..3628
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3674..4102
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4116..4445
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4512..5306
                     /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    5480..5601
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5650..5666)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5674..5690
                     /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(5698..5728)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5743..5764
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6052..6637)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6808..7668)
                     /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(7669..7773)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgcggccg gagcccggag gctgctgctg ccgccgcacg
     1681 gtgcgggcga atggctgcgt ggcgaacggg gaagtacgga acgggtacgt gaggagcagc
     1741 gctgcagccg cagccgcagc cgccgccggc cagatccatc atgttacaca aaatggagga
     1801 ctatataaaa gaccgtttaa tgaagctttt gaagaaacac caatgctggt tgctgtgctc
     1861 acgtatgtgg ggtatggcgt actcaccctc tttggatatc ttcgagattt cttgaggtat
     1921 tggagaattg aaaagtgtca ccatgcaaca gaaagagaag aacaaaagga ctttgtgtca
     1981 ttgtatcaag attttgaaaa cttttataca aggaatctgt acatgaggat aagagacaac
     2041 tggaatcggc caatctgtag tgtgcctgga gccagggtgg acatcatgga gagacagtct
     2101 catgattata actggtcctt caagtataca gggaatataa taaagggtgt tataaacatg
     2161 ggttcctaca actatcttgg atttgcacgg aatactggat catgtcaaga agcagccgcc
     2221 aaagtccttg aggagtatgg agctggagtg tgcagtactc ggcaggaaat tggaaacctg
     2281 gacaagcatg aagaactaga ggagcttgta gcaaggttct taggagtaga agctgctatg
     2341 gcgtatggca tgggatttgc aacgaattca atgaacattc ctgctcttgt tggcaaaggt
     2401 tgcctgattc tgagtgatga actgaatcat gcatcactgg ttctgggagc cagactgtca
     2461 ggagcaacca ttagaatctt caaacacaac aatatgcaaa gcctagagaa gctattgaaa
     2521 gatgccattg tttatggtca gcctcggaca cgaaggccct ggaagaaaat tctcatcctt
     2581 gtggaaggaa tatatagcat ggagggatct attgttcgtc ttcctgaagt gattgccctc
     2641 aagaagaaat acaaggcata cttgtatctg gatgaggctc acagcattgg cgccctgggc
     2701 cccacaggcc ggggtgtggt ggagtacttt ggcctggatc ccgaggatgt ggatgttatg
     2761 atgggaacgt tcacaaagag ttttggtgct tctggaggat atattggagg caagaaggag
     2821 ctgatagact acctgcgaac acattctcat agtgcagtgt atgccacgtc attgtcacct
     2881 cctgtagtgg agcagatcat cacctccatg aagtgcatca tggggcagga tggcaccagc
     2941 cttggtaaag agtgtgtaca acagttagct gaaaacacca ggtatttcag gagacgcctg
     3001 aaagagatgg gcttcatcat ctatggaaat gaagactctc cagtagtgcc tttgatgctc
     3061 tacatgcctg ccaaaattgg cgcctttgga cgggagatgc tgaagcggaa catcggtgtc
     3121 gttgtggttg gatttcctgc caccccaatt attgagtcca gagccaggtt ttgcctgtca
     3181 gcagctcata ccaaagaaat acttgatact gctttaaagg agatagatga agttggggac
     3241 ctattgcagc tgaagtattc ccgtcatcgg ttggtacctc tactggacag gccctttgac
     3301 gagacgacgt atgaagaaac agaagactga ggatcctgat atccagcaca gtggcggccg
     3361 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3421 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3481 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3541 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3601 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3661 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3721 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3781 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3841 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3901 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3961 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4021 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4081 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4321 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4381 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4441 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4501 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4561 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4621 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4681 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4741 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4801 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4861 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4921 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4981 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5041 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5101 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5161 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5221 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5281 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5341 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5401 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5461 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5521 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5581 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5641 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5701 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5761 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5821 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5881 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5941 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6241 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6481 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6781 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6841 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6901 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6961 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7021 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7081 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7141 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7201 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7261 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7321 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7381 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7441 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7741 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7801 cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
