LOCUS       V041758                 7980 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041758
VERSION     V041758
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..2783
                     /label="RHOT2(NM_001352275)"
                     /note="RHOT2(NM_001352275)"
                     /gene="RHOT2"
     CDS             2823..3542
                     /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    3580..3804
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3850..4278
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4292..4621
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4688..5482
                     /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    5656..5777
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5826..5842)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5850..5866
                     /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(5874..5904)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5919..5940
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6228..6813)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6984..7844)
                     /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(7845..7949)
                     /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 gccaccatga ggcgggacgt gcgcatcctg ttactgggcg
      961 aggcccaggt ggggaagacg tcgctgatcc tgtccctggt gggcgaggag ttccccgagg
     1021 aggtccctcc ccgcgcggag gagatcacca tccccgcgga cgtcaccccg gagaaggtgc
     1081 ccacccacat cgtggactac tcagaagccg agcagacgga cgaggagctg cgggaggaga
     1141 tccacaaggc aaacgtggtg tgtgtggtgt atgacgtctc tgaggaggcc accattgaga
     1201 agattcgaac taagtggatc ccactggtga atggggggac cacgcagggg cccagggtgc
     1261 ccatcatcct agtgggcaac aagtcagacc tgcggtcggg gagctccatg gaggccgtgc
     1321 tccccatcat gagccagttt cccgagattg agacctgcgt ggagtgttcg gccaagaacc
     1381 tgaggaacat ctcagagctg ttctactacg cccagaaggc cgtcctgcat cccacagccc
     1441 ccctctatga ccctgaggcc aagcagttga ggcccgcgtg cgcccaggcg ctgacgcgca
     1501 tcttcaggct ctcagatcag gacctggacc aggcgctcag tgacgaagag ctcaacgctt
     1561 tccagcagaa atcctgcttt gggcaccccc tggccccgca ggccctggag gacgtgaaga
     1621 cggtggtgtg caggaacgtg gcgggcggcg tgcgggagga ccggctgacc ctggatggtt
     1681 tcctcttcct gaacacgctc ttcatccagc gcggccggca cgagaccacc tggaccatcc
     1741 tgcggcgctt cggctacagc gatgccctgg agctgactgc ggactatctc tcccctctga
     1801 tccacgtgcc ccccggctgc agcacggagc tcaaccacct tggctaccag tttgtgcaga
     1861 gagtgtttga gaagcacgac caggaccgcg acggcgccct ctcgcccgtg gagctgcaaa
     1921 gccttttcag tgtgttccca gcagcgccct ggggccccga gctcccacgc acagtccgca
     1981 cagaggccgg ccggttgccc ctgcacggat acctctgcca gtggaccctg gtgacctacc
     2041 tggacgtccg gagctgcctt ggacacctag gctacctggg ctaccccacc ctctgtgagc
     2101 aggaccaggc ccatgccatc acagtcactc gtgagaagag gctggaccag gagaagggac
     2161 agacgcagcg gagcgtcctc ctgtgcaagg tggtaggggc ccgtggagtg ggcaagtctg
     2221 ccttcctgca ggcctttctc ggccgcggcc tggggcacca ggacacgagg gagcagcctc
     2281 ccggctacgc catcgacacg gtgcaggtca atggacagga gaagtacttg atcctctgtg
     2341 aggtgggcac agatggtctg ctggccacat cgctggacgc cacctgtgac gttgcctgct
     2401 tgatgtttga tggcagtgac ccaaagtcct ttgcacattg tgccagcgtc tacaagcacc
     2461 attacatgga cgggcagacc ccctgcctct ttgtctcctc caaggccgac ctgcccgaag
     2521 gtgtcgcggt gtctggccca tcaccggccg agttttgccg caagcaccgg ctacccgctc
     2581 ccgtgccgtt ctcctgtgct ggcccagccg agcccagcac caccatcttc acccagctcg
     2641 ccaccatggc cgccttccca catttggtcc acgcagagct gcatccctct tccttctggc
     2701 tccgggggct gctgggggtt gtcggggccg ccgtggccgc agtcctcagc ttctcactct
     2761 acagggtcct ggtgaagagc caggcctcga gtggcggtgg aggaagcggc ggaggtggat
     2821 ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2881 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2941 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3001 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3061 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3121 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3181 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3241 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3301 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3361 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3421 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3481 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3541 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3601 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3661 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3721 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3781 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3841 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3901 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3961 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4021 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4081 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4141 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4201 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4261 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4441 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4501 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4561 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4621 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4681 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4741 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4801 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4861 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4921 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4981 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5041 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5101 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5161 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5221 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5281 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5341 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5401 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5461 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5521 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5581 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5641 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5761 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5821 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5881 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5941 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6001 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6061 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6121 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6181 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6241 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6301 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6361 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6421 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6481 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6541 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6601 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6661 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6721 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     6781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
