LOCUS       V036124                 7512 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036124
VERSION     V036124
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             927..2315
                     /label="GLP1R(NM_002062)"
                     /note="GLP1R(NM_002062)"
                     /gene="GLP1R"
     CDS             2355..3074
                     /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    3112..3336
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3382..3810
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3824..4153
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4220..5014
                     /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    5188..5309
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5358..5374)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5382..5398
                     /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(5406..5436)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5451..5472
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5760..6345)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6516..7376)
                     /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(7377..7481)
                     /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 gtttaaagcc actgggatcc gccaccatgg ccggcgcccc cggcccgctg cgccttgcgc
      961 tgctgctgct cgggatggtg ggcagggccg gcccccgccc ccagggtgcc actgtgtccc
     1021 tctgggagac ggtgcagaaa tggcgagaat accgacgcca gtgccagcgc tccctgactg
     1081 aggatccacc tcctgccaca gacttgttct gcaaccggac cttcgatgaa tacgcctgct
     1141 ggccagatgg ggagccaggc tcgttcgtga atgtcagctg cccctggtac ctgccctggg
     1201 ccagcagtgt gccgcagggc cacgtgtacc ggttctgcac agctgaaggc ctctggctgc
     1261 agaaggacaa ctccagcctg ccctggaggg acttgtcgga gtgcgaggag tccaagcgag
     1321 gggaaagaag ctccccggag gagcagctcc tgttcctcta catcatctac acggtgggct
     1381 acgcactctc cttctctgct ctggttatcg cctctgcgat cctcctcggc ttcagacacc
     1441 tgcactgcac caggaactac atccacctga acctgtttgc atccttcatc ctgcgagcat
     1501 tgtccgtctt catcaaggac gcagccctga agtggatgta tagcacagcc gcccagcagc
     1561 accagtggga tgggctcctc tcctaccagg actctctgag ctgccgcctg gtgtttctgc
     1621 tcatgcagta ctgtgtggcg gccaattact actggctctt ggtggagggc gtgtacctgt
     1681 acacactgct ggccttctcg gtcttatctg agcaatggat cttcaggctc tacgtgagca
     1741 taggctgggg tgttcccctg ctgtttgttg tcccctgggg cattgtcaag tacctctatg
     1801 aggacgaggg ctgctggacc aggaactcca acatgaacta ctggctcatt atccggctgc
     1861 ccattctctt tgccattggg gtgaacttcc tcatctttgt tcgggtcatc tgcatcgtgg
     1921 tatccaaact gaaggccaat ctcatgtgca agacagacat caaatgcaga cttgccaagt
     1981 ccacgctgac actcatcccc ctgctgggga ctcatgaggt catctttgcc tttgtgatgg
     2041 acgagcacgc ccgggggacc ctgcgcttca tcaagctgtt tacagagctc tccttcacct
     2101 ccttccaggg gctgatggtg gccatattat actgctttgt caacaatgag gtccagctgg
     2161 aatttcggaa gagctgggag cgctggcggc ttgagcactt gcacatccag agggacagca
     2221 gcatgaagcc cctcaagtgt cccaccagca gcctgagcag tggagccacg gcgggcagca
     2281 gcatgtacac agccacttgc caggcctcct gcagctgctc gagtggcggt ggaggaagcg
     2341 gcggaggtgg atctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2401 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2461 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2521 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2581 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2641 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2701 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2761 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2821 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2881 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2941 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3001 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3061 agctgtacaa gtagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     3121 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3181 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3241 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3301 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3361 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3421 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3481 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3541 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3601 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3661 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3721 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3781 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3961 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4021 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4081 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4141 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4201 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4261 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4321 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4381 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4441 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4501 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4561 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4621 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4681 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4741 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4801 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4861 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4921 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4981 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5041 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5101 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5161 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5221 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5281 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5341 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7501 ccacctgacg tc
//
