LOCUS       V038144                 8068 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038144
VERSION     V038144
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..3558
                     /label="PARN(NM_002582)"
                     /note="PARN(NM_002582)"
                     /gene="PARN"
     polyA_signal    3635..3859
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3905..4333
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4347..4676
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4743..5537
                     /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    5711..5832
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5881..5897)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5905..5921
                     /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(5929..5959)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5974..5995
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6283..6868)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7039..7899)
                     /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(7900..8004)
                     /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 catggagata atcaggagca attttaagag taatcttcac
     1681 aaagtgtacc aggccataga ggaggccgac ttcttcgcca tcgatgggga gttttcagga
     1741 atcagtgatg gaccttcagt ctctgcatta acaaatggtt ttgacactcc agaagagagg
     1801 tatcagaagc ttaaaaagca ttccatggac tttttgctat ttcagtttgg cctttgcact
     1861 tttaagtatg actacacaga ttcaaagtat ataacgaagt catttaactt ctatgttttc
     1921 ccgaaaccct tcaatagatc ctcaccagat gtcaaatttg tttgtcagag ctccagcatt
     1981 gactttctag caagccaggg atttgatttt aataaagttt ttcgaaatgg aattccatat
     2041 ttaaatcagg aagaagaaag acagttaaga gagcagtatg atgaaaaacg ttcacaggcg
     2101 aatggtgcag gagctctgtc ctatgtatct cctaacactt caaaatgtcc tgtcacgatt
     2161 cctgaggatc aaaagaagtt tattgaccaa gtggtagaga aaatagagga tttattacaa
     2221 agtgaagaaa acaagaactt ggatttagag ccatgtaccg ggttccaaag aaaactaatt
     2281 tatcagactt tgagctggaa gtatccgaaa ggcattcatg ttgagacttt agaaactgaa
     2341 aagaaggagc gatatatagt tatcagcaaa gtagatgaag aagaacgcaa aagaagagag
     2401 cagcagaaac atgccaaaga acaggaggag ctgaatgatg ctgtgggatt ttctagagtc
     2461 attcacgcca ttgctaattc gggaaaactt gttattggac acaatatgct cttggacgtc
     2521 atgcacacag ttcatcagtt ctactgccct ctgcctgcgg acttaagtga gtttaaagag
     2581 atgacaacat gtgttttccc cagactcttg gatactaaat tgatggccag cacacaacct
     2641 tttaaggata tcattaacaa cacatccctt gcggaattgg aaaagcggtt aaaagagaca
     2701 cctttcaacc ctcctaaagt tgaaagtgcc gaaggttttc caagttatga cacagcctct
     2761 gaacaactcc acgaggcagg ctacgatgcc tacatcacag ggctgtgctt catctccatg
     2821 gccaattacc taggttcttt tctcagccct ccaaaaattc atgtgtctgc cagatcaaaa
     2881 ctcattgaac ctttttttaa caagttattt cttatgaggg tcatggatat cccctatcta
     2941 aacttggaag gaccagactt gcagcctaaa cgtgatcatg ttctccatgt gacattcccc
     3001 aaagaatgga aaaccagcga cctttaccag cttttcagtg cctttggtaa cattcagata
     3061 tcctggattg atgacacatc agcatttgtt tcccttagcc agcccgagca agtaaagatt
     3121 gctgtcaata ccagcaaata tgcagaaagc tatcggatcc aaacctatgc tgaatatatg
     3181 gggagaaaac aggaagagaa gcagatcaaa agaaagtgga ctgaagatag ctggaaggag
     3241 gctgacagca aacggttaaa cccccagtgc ataccctaca ccctgcagaa tcactattac
     3301 cgcaacaata gttttacagc tcccagcaca gtaggaaaga gaaatttgag tcctagtcaa
     3361 gaggaagctg gcctggagga cggagtgtca ggggagattt ccgacactga gcttgagcag
     3421 accgattcct gtgcagagcc cctctcagag ggaaggaaaa aggccaagaa attaaaaaga
     3481 atgaagaagg agctttctcc agcaggaagc atctcgaaga acagccctgc cacactcttt
     3541 gaagttcctg acacatggta aggatcctga tatccagcac agtggcggcc gctcgagtct
     3601 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3661 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3721 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3781 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3841 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3901 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3961 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4021 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4081 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4141 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4201 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4261 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4321 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4561 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4621 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4681 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4741 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4801 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4861 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4921 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4981 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5041 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5101 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5161 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5221 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5281 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5341 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5401 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5461 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5521 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5581 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5641 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5701 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5821 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5881 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5941 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6001 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8041 atcgggagat ctcccgatcc cctatggt
//
