LOCUS       V040524                 6697 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040524
VERSION     V040524
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..2133
                     /label="OPN3(NM_014322)"
                     /note="OPN3(NM_014322)"
                     /gene="OPN3"
     CDS             2140..2163
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2170..2259
                     /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    2297..2521
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2567..2995
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3009..3338
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3405..4199
                     /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    4373..4494
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4543..4559)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4567..4583
                     /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(4591..4621)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4636..4657
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4945..5530)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5701..6561)
                     /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(6562..6666)
                     /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 tactcgggga accgcagcgg cggccacggc
      961 tactgggacg gcggcggggc cgcgggcgct gaggggccgg cgccggcggg gacactgagc
     1021 cccgcgcccc tcttcagccc cggcacctac gagcgcctgg cgctgctgct gggctccatt
     1081 gggctgctgg gcgtcggcaa caacctgctg gtgctcgtcc tctactacaa gttccagcgg
     1141 ctccgcactc ccactcacct cctcctggtc aacatcagcc tcagcgacct gctggtgtcc
     1201 ctcttcgggg tcacctttac cttcgtgtcc tgcctgagga acggctgggt gtgggacacc
     1261 gtgggctgcg tgtgggacgg gtttagcggc agcctcttcg ggattgtttc cattgccacc
     1321 ctaaccgtgc tggcctatga acgttacatt cgcgtggtcc atgccagagt gatcaatttt
     1381 tcctgggcct ggagggccat tacctacatc tggctctact cactggcgtg ggcaggagca
     1441 cctctcctgg gatggaacag gtacatcctg gacgtacacg gactaggctg cactgtggac
     1501 tggaaatcca aggatgccaa cgattcctcc tttgtgcttt tcttatttct tggctgcctg
     1561 gtggtgcccc tgggtgtcat agcccattgc tatggccata ttctatattc cattcgaatg
     1621 cttcgttgtg tggaagatct tcagacaatt caagtgatca agattttaaa atatgaaaag
     1681 aaactggcca aaatgtgctt tttaatgata ttcaccttcc tggtctgttg gatgccttat
     1741 atcgtgatct gcttcttggt ggttaatggt catggtcacc tggtcactcc aacaatatct
     1801 attgtttcgt acctctttgc taaatcgaac actgtataca atccagtgat ttatgtcttc
     1861 atgatcagaa agtttcgaag atcccttttg cagcttctgt gcctccgact gctgaggtgc
     1921 cagaggcctg ctaaagacct accagcagct ggaagtgaaa tgcagatcag acccattgtg
     1981 atgtcacaga aagatgggga caggccaaag aaaaaagtga ctttcaactc ttcttccatc
     2041 atttttatca tcaccagtga tgaatcactg tcagttgacg acagcgacaa aaccaatggg
     2101 tccaaagttg atgtaatcca agttcgtcct ttgctcgagg attacaagga tgacgacgat
     2161 aagggatcca gcgcctggtc ccaccctcag tttgagaagg gcggaggctc tggcggcgga
     2221 agcggaggat ctgcttggag ccacccccag ttcgaaaagt gatgagggcc cgtttaaacc
     2281 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2341 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2401 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2461 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2521 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2641 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2701 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2761 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2821 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2881 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2941 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3001 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3241 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3301 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3361 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3421 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3481 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3541 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3601 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3661 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3721 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3781 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3841 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3901 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3961 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4021 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4081 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4141 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4201 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4261 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4321 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4501 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4561 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4621 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4681 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4741 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4801 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4861 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5461 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5761 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5821 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5881 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5941 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6001 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6061 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6121 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6181 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6241 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6301 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6361 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6421 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6481 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6541 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6601 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6661 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
