LOCUS       V033135                 7525 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033135
VERSION     V033135
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..3015
                     /label="Tmem135(NM_028343)"
                     /note="Tmem135(NM_028343)"
                     /gene="Tmem135"
     polyA_signal    3092..3316
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3362..3790
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3804..4133
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4200..4994
                     /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    5168..5289
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5338..5354)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5362..5378
                     /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(5386..5416)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5431..5452
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5740..6325)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6496..7356)
                     /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(7357..7461)
                     /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 catggcggcc ctcagcaagt ccatccctca taactgctac
     1681 gagatcggcc acacttggca cccctcctgc agggtctcct tcctgcagat cacttggggc
     1741 gccctggagg agtccctcag gatctacgca cccctgtacc tgattgcagc tgttctccgg
     1801 aagcgcaagc tggagtatta cctgtacaaa ctgcttcctg agatcctcca gtccgcctcc
     1861 ttcctgactg ctaatggggc cttgtacatt actttcttct gcattttaag gaagatcctt
     1921 ggaaaattct actcatggac tcctggcttt ggtgctgctc tgccagcctc atatgtggcc
     1981 attctaattg aaagaaaaag caggagaggg ctgctcacaa tttatatggc caacttggcc
     2041 acagaaacac tatttagaat gggtgttgca agaggaacca tcacgacgtt aagaaatgga
     2101 gaagtccttt tgttctgcat cacagcggcc atgtacatgt tctttttcag gtgcaaagat
     2161 ggcttaaaag gctttacatt ttctgcactt aggtttatcg tggggaagga agaaattccc
     2221 acgcactctt actcaccaga gacagcgtat gccaaagtgg aacagaagag ggagaagcac
     2281 aagggaacgc cgagagcaat gagcataatc gctctggttc ggactctcgt ggactccgta
     2341 tgcaagcatg gaccaaggca cagatgctgc aagcattatg aagacaactg catttcctac
     2401 tgcattaaag gcttcatcag aatgttcagc gtgggctact tgatccagtg ctgtctccgg
     2461 atcccctctg cgtttaggca tctgtttaca gaaccatccc ggctcctctc tcttttctac
     2521 aacaaggaga acttccagct gggggctttc cttggatctt ttgttagtat atacaagggc
     2581 acaagttgtt tcctgcgctg gatcaggaat ctggatgacg agctgcatgc tattgtggct
     2641 gggtttttgg caggtgtgtc gatgatgttt tataaaagca caacaatttc catgtaccta
     2701 gcttccaagc tggtggagac aatgtatttc aagggcatcg aggctggaaa ggttccctat
     2761 tttcctcaag cagatacgat catctattcc atctctacag caatttgttt ccacgcagct
     2821 gttatggaag ttcagaactt gcgaccatct tactggaagt tccttttaag gctcaccaag
     2881 ggcaggtttg cactcatgaa ccgaaaagcc ctggatgtgt ttggtactgg cgcatctaga
     2941 gagtttcaca acttcatccc cagactagac ccacggtaca cagttgtcac accggagctg
     3001 cccatagact tttcctgagg atcctgatat ccagcacagt ggcggccgct cgagtctaga
     3061 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3121 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3181 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3241 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3301 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3361 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3421 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3481 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3541 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3601 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3661 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3721 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3781 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4081 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4141 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4201 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4261 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4321 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4381 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4441 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4501 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4561 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4621 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4681 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4741 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4801 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4861 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4921 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4981 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5041 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5101 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5161 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5281 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7501 gggagatctc ccgatcccct atggt
//
