LOCUS       V030572                 8335 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030572
VERSION     V030572
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             931..2730
                     /label="GPR107(NM_001136557)"
                     /note="GPR107(NM_001136557)"
                     /gene="GPR107"
     CDS             2857..3858
                     /codon_start=1
                     /note="In the Tet-Off® system, tTA binds to the
                     Tet-responsive element&nbsp;and stimulates transcription
                     only in the absence of tetracycline or doxycycline."
                     /product="tetracycline-controlled transactivator,
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus
                     VP16"
                     /transl_table=1
                     /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN
                     KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT
                     EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS
                     MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS,AYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG*"
                     /label="tTA"
     polyA_signal    3902..4126
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4172..4600
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4614..4943
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5010..5804
                     /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    5978..6099
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6148..6164)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6172..6188
                     /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(6196..6226)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6241..6262
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6550..7135)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7306..8166)
                     /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(8167..8271)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggccgctc tggcgcccgt cggctccccc
      961 gcctcccgcg gtcctaggct ggccgcgggc ctccggctgc tcccaatgct gggtttgctg
     1021 cagttgctgg ccgagcctgg cctgggccgc gtccatcacc tggcactcaa ggatgatgtg
     1081 aggcataaag ttcatctgaa cacctttggc ttcttcaagg atgggtacat ggtggtgaat
     1141 gtcagtagcc tctcactgaa tgagcctgaa gacaaggatg tgactattgg atttagccta
     1201 gaccgtacaa agaatgatgg cttttcttct tacctggatg aagatgtgaa ttactgtatt
     1261 ttaaagaaac agtctgtctc tgtcaccctt ttaatcctag acatctccag aagtgaggta
     1321 agagtaaagt ctccaccaga agctggtacc cagttaccaa agatcatctt cagcagggat
     1381 gagaaagtcc ttggtcagag ccaggagcct aatgttaacc ctgcttcagc aggcaaccag
     1441 acccagaaga cacaagatgg tggaaagtct aaaagaagta cagtggattc aaaggccatg
     1501 ggagagaaat ccttttctgt tcataataat ggtggggcag tgtcatttca gtttttcttt
     1561 aacatcagca ctgatgacca agaaggcctt tacagtcttt attttcataa atgccttgga
     1621 aaagaattgc caagtgacaa gtttacattc agccttgata ttgagatcac agagaagaat
     1681 cctgacagct acctctcagc aggagaaatt cctctcccca aattatacat ctcaatggcc
     1741 tttttcttct ttctttctgg gaccatctgg attcatatcc ttcgaaaacg acggaatgat
     1801 gtatttaaaa tccactggct gatggcggcc cttcctttca ccaagtctct ttccttggtg
     1861 ttccatgcaa ttgactacca ctacatctcc tcccagggct tccctatcga aggctgggct
     1921 gttgtgtact acataactca ccttttgaaa ggggcgctac tcttcatcac cattgcactc
     1981 attggcactg gctgggcttt cattaagcac atcctttctg ataaagacaa aaagatcttc
     2041 atgattgtca ttccactcca ggtcctggca aatgtagcct acatcatcat agagtccacc
     2101 gaggagggca cgactgaata tggcttgtgg aaggactctc tatttctggt cgacctgttg
     2161 tgttgtggtg ccatcctctt cccagtggtg tggtcaatca gacatttaca agaagcatca
     2221 gcaacagatg gaaaaggtga cagcatggga cctcttcagc agagagcgaa tctaagagca
     2281 ggaagtcgca tagagtctca ccattttgcc caggctgatc ttgaactcct ggcctctagc
     2341 tgtcctcctg cctcagtctc ccaaagggct gggattacag ctgctattaa cttagcaaag
     2401 ctgaaacttt tcagacatta ttacgtcttg attgtgtgtt acatatactt cactaggatc
     2461 attgcatttc tcctcaaact cgctgttcca ttccagtgga agtggctcta ccagctcctg
     2521 gatgaaacgg ccacactggt cttctttgtt ctaacggggt ataaattccg tccggcttca
     2581 gataacccct acctacaact ttctcaggaa gaagaagact tggaaatgga gtccgttgtg
     2641 acaacatctg gggtgatgga aagtatgaag aaagtcaaga aggtgaccaa cggctccgtg
     2701 gagccccagg gcgagtggga aggcgccgtg gatatcatcg ataccggtgg acgcacccca
     2761 cccagcctgg gtccccaaga tgagtcctgc accaccgcca gctcctccct ggccaaggac
     2821 acttcatcga ccggtgagaa cctgtacttc cagctaagat tagataaaag taaagtgatt
     2881 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     2941 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     3001 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     3061 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     3121 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     3181 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     3241 ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag
     3301 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     3361 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     3421 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc cgcgtacagc
     3481 cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga tctcccggac
     3541 gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct ccccgcggga
     3601 cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga cgagctccac
     3661 ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt cgatctggac
     3721 atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc cgccccctac
     3781 ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc ccttggaatt
     3841 gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3901 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3961 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4021 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4081 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4141 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4201 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4261 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4321 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4381 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4441 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4501 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4561 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4621 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4681 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4741 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4801 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4861 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4921 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4981 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5041 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5101 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5161 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5221 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5281 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5341 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5401 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5461 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5521 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5581 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5641 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5701 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5761 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5821 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5881 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5941 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6001 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6061 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6121 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6181 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6241 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6301 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6361 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7081 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7141 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7201 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7261 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7321 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7381 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7441 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7501 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7561 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7621 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7681 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7741 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7801 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7861 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7921 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7981 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8041 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8101 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8161 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8221 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8281 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
