LOCUS       V021448                 7771 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021448
VERSION     V021448
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             649..1365
                     /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             1399..2481
                     /label="LSAMP(NM_001318915)"
                     /note="LSAMP(NM_001318915)"
                     /gene="LSAMP"
     misc_feature    2547..3135
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3201..3249
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3451..3879
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3893..4222
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4289..5083
                     /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    5259..5380
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5429..5445)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5453..5469
                     /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(5477..5507)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5522..5543
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5831..6419)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6590..7450)
                     /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(7451..7555)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat ggtcaggaga gttcagccgg atcggaaaca gttgccactg
     1441 gtcctactga gattgctctg ccttcttccc acaggactgc ctgttcgcag cgtggatttt
     1501 aaccgaggca cggacaacat caccgtgagg cagggggaca cagccatcct caggtgcgtt
     1561 gtagaagaca agaactcaaa ggtggcctgg ttgaaccgtt ctggcatcat ttttgctgga
     1621 catgacaagt ggtctctgga cccacgggtt gagctggaga aacgccattc tctggaatac
     1681 agcctccgaa tccagaaggt ggatgtctat gatgagggtt cctacacttg ctcagttcag
     1741 acacagcatg agcccaagac ctcccaagtt tacttgatcg tacaagtccc accaaagatc
     1801 tccaatatct cctcggatgt cactgtgaat gagggcagca acgtgactct ggtctgcatg
     1861 gccaatggcc gtcctgaacc tgttatcacc tggagacacc ttacaccaac tggaagggaa
     1921 tttgaaggag aagaagaata tctggagatc cttggcatca ccagggagca gtcaggcaaa
     1981 tatgagtgca aagctgccaa cgaggtctcc tcggcggatg tcaaacaagt caaggtcact
     2041 gtgaactatc ctcccactat cacagaatcc aagagcaatg aagccaccac aggacgacaa
     2101 gcttcactca aatgtgaggc ctcggcagtg cctgcacctg actttgagtg gtaccgggat
     2161 gacactagga taaatagtgc caatggcctt gagattaaga gcacggaggg ccagtcttcc
     2221 ctgacggtga ccaacgtcac tgaggagcac tacggcaact acacctgtgt ggctgccaac
     2281 aagctggggg tcaccaatgc cagcctagtc cttttcaaac gtgttttacc cacaatcccc
     2341 caccccattc aagaaattgg taccaccgtg cacttcaagc aaaaaggacc tgggtcggtg
     2401 agaggaataa atggatccat cagtctggcc gtaccactgt ggctgctggc agcatctctg
     2461 ctctgccttc tcagcaaatg ttaagaattc tgcagatatc cagcacagtg gcggccgctc
     2521 gagtctagag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2581 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2641 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2701 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2761 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2821 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2881 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2941 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3001 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3061 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3121 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     3181 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     3241 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3301 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3361 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3421 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3481 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3541 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3601 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3661 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3721 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3781 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3841 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3901 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5161 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5221 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5401 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5461 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5581 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5641 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5701 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5761 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5821 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5881 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5941 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6001 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6061 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6241 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6361 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6481 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6541 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6601 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6661 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6721 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6781 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6841 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6901 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6961 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7021 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7081 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7141 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7201 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7261 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7321 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7381 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7441 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7501 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7561 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7621 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7681 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     7741 gaccgacaat tgcatgaaga atctgcttag g
//
