LOCUS       V040025                 6535 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040025
VERSION     V040025
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             928..1971
                     /label="RHO(NM_000539)"
                     /note="RHO(NM_000539)"
                     /gene="RHO"
     CDS             1978..2001
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2008..2097
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2135..2359
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2405..2833
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2847..3176
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3243..4037
                     /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    4211..4332
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4381..4397)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4405..4421
                     /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(4429..4459)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4474..4495
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4783..5368)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5539..6399)
                     /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(6400..6504)
                     /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 gtttaaactt aagctgaatt cgccaccatg aatggcacag aaggccctaa cttctacgtg
      961 cccttctcca atgcgacggg tgtggtacgc agccccttcg agtacccaca gtactacctg
     1021 gctgagccat ggcagttctc catgctggcc gcctacatgt ttctgctgat cgtgctgggc
     1081 ttccccatca acttcctcac gctctacgtc accgtccagc acaagaagct gcgcacgcct
     1141 ctcaactaca tcctgctcaa cctagccgtg gctgacctct tcatggtcct aggtggcttc
     1201 accagcaccc tctacacctc tctgcatgga tacttcgtct tcgggcccac aggatgcaat
     1261 ttggagggct tctttgccac cctgggcggt gaaattgccc tgtggtcctt ggtggtcctg
     1321 gccatcgagc ggtacgtggt ggtgtgtaag cccatgagca acttccgctt cggggagaac
     1381 catgccatca tgggcgttgc cttcacctgg gtcatggcgc tggcctgcgc cgcaccccca
     1441 ctcgccggct ggtccaggta catccccgag ggcctgcagt gctcgtgtgg aatcgactac
     1501 tacacgctca agccggaggt caacaacgag tcttttgtca tctacatgtt cgtggtccac
     1561 ttcaccatcc ccatgattat catctttttc tgctatgggc agctcgtctt caccgtcaag
     1621 gaggccgctg cccagcagca ggagtcagcc accacacaga aggcagagaa ggaggtcacc
     1681 cgcatggtca tcatcatggt catcgctttc ctgatctgct gggtgcccta cgccagcgtg
     1741 gcattctaca tcttcaccca ccagggctcc aacttcggtc ccatcttcat gaccatccca
     1801 gcgttctttg ccaagagcgc cgccatctac aaccctgtca tctatatcat gatgaacaag
     1861 cagttccgga actgcatgct caccaccatc tgctgcggca agaacccact gggtgacgat
     1921 gaggcctctg ctaccgtgtc caagacggag acgagccagg tggccccggc cctcgaggat
     1981 tacaaggatg acgacgataa gggatccagc gcctggtccc accctcagtt tgagaagggc
     2041 ggaggctctg gcggcggaag cggaggatct gcttggagcc acccccagtt cgaaaagtga
     2101 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2161 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2221 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2281 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2341 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2401 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2461 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2821 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2881 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2941 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3001 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3061 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3121 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3181 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3241 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3301 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3361 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3421 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3481 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3541 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3601 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3661 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3721 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3781 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3841 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3901 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3961 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4021 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4081 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4201 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4261 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4321 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     5341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5461 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5521 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5581 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5641 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5701 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5761 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5821 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5881 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5941 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6001 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6061 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6121 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6181 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6241 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6301 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6361 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6421 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6481 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
