LOCUS       V035945                 7761 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035945
VERSION     V035945
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             709..726
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
     CDS             730..2487
                     /label="CRY1(NM_004075)"
                     /note="CRY1(NM_004075)"
                     /gene="CRY1"
     CDS             2494..2517
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2537..3125
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3191..3239
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3441..3869
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3883..4212
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4279..5073
                     /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    5249..5370
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5419..5435)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5443..5459
                     /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(5467..5497)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5512..5533
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5821..6409)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6580..7440)
                     /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(7441..7545)
                     /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 gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgct ggtgccgcgc
      721 ggcagccata tgggggtgaa cgccgtgcac tggttccgaa aggggctccg gctccacgac
      781 aaccccgccc tgaaggagtg cattcagggc gccgacacca tccgctgcgt ctacatcctg
      841 gacccctggt tcgccggctc ctccaatgtg ggcatcaaca ggtggcgatt tttgcttcag
      901 tgtcttgagg atcttgatgc caatctacga aaattaaact cccgtctgtt tgtgattcgt
      961 ggacaaccag cagatgtgtt tcccaggctt ttcaaggaat ggaacattac taaactttca
     1021 attgagtatg attctgagcc ctttggaaag gaacgagacg cagctattaa gaaactggca
     1081 actgaagctg gagtagaagt cattgtaaga atttcacata cattatatga cctagacaag
     1141 atcatagaac tcaatggtgg acaaccgcct ctaacttata aaagattcca gactctcatc
     1201 agcaaaatgg aaccactaga gataccagta gagacaatta cttcagaagt gatagaaaag
     1261 tgcacaactc ctctgtctga tgaccatgat gagaaatatg gagtcccttc actggaagag
     1321 ctaggttttg atacagatgg cttatcctct gcagtgtggc caggtggaga aactgaagca
     1381 cttactcgtt tggaaaggca tttggaaaga aaagcttggg tggcaaattt tgaaagacct
     1441 cgaatgaatg cgaattctct gcttgcaagc cctactggac ttagtcctta tctccgattt
     1501 ggttgtttgt catgtcgact gttttacttc aaactaacag atctctacaa aaaggtaaag
     1561 aagaacagtt cccctcccct ttccctttat gggcaactgt tatggcgtga atttttctat
     1621 acagcagcaa caaataatcc acgctttgat aaaatggaag gaaaccctat ctgtgttcag
     1681 attccttggg ataaaaatcc tgaggcttta gccaaatggg cggaaggccg gacaggcttt
     1741 ccatggattg atgccatcat gacacagctt cgtcaggagg gttggattca tcatctagcc
     1801 aggcatgcag ttgcttgctt cctgacacga ggggacctgt ggattagttg ggaagaagga
     1861 atgaaggtat ttgaagaatt attgcttgat gcagattgga gcataaatgc tggaagttgg
     1921 atgtggctgt cttgtagttc cttttttcaa cagttttttc actgctattg ccctgttggt
     1981 tttggtagga gaacagatcc caatggagac tatatcaggc gttatttgcc tgtcctaaga
     2041 ggcttccctg caaaatatat ctatgatccc tggaatgcac cagaaggtat ccaaaaggta
     2101 gccaaatgtt tgataggagt taattatcct aaaccaatgg tgaaccatgc tgaggcaagc
     2161 cgtttgaata tcgaaaggat gaaacagatc tatcagcagc tttcacgata tagaggacta
     2221 ggtcttctgg catcagtacc ttctaatcct aatgggaatg gaggcttcat gggatattct
     2281 gcagaaaata tcccaggttg tagcagcagt ggaagttgct ctcaagggag tggtatttta
     2341 cactatgctc atggcgacag tcagcaaact cacctgttga agcaaggaag aagctccatg
     2401 ggcactggtc tcagtggtgg gaaacgtcct agtcaggaag aggacacaca gagtattggt
     2461 cctaaagtcc agagacagag cactaatctc gagcaccatc accatcacca tcaccattag
     2521 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2581 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2641 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2701 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2761 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2821 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2881 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     2941 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3001 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3061 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3121 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3181 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     3241 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3301 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3361 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     3421 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3661 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3721 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3781 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3841 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4261 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4321 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4381 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4441 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4501 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4561 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4621 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4681 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4741 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4801 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4861 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4921 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4981 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5041 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     5101 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5161 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5221 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5341 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5401 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5461 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6001 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6241 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7621 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7681 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7741 tgcatgaaga atctgcttag g
//
