LOCUS       V024358                 9252 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024358
VERSION     V024358
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1500..3932
                     /label="OSBPL5(NM_145638)"
                     /note="OSBPL5(NM_145638)"
                     /gene="OSBPL5"
     CDS             3939..4004
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4028..4616
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4682..4730
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4932..5360
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5374..5703
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5770..6564
                     /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    6740..6861
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6910..6926)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6934..6950
                     /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(6958..6988)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7003..7024
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7312..7900)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8071..8931)
                     /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(8932..9036)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tgaaggagga ggccttcctc cggcgccgct tctccctgtg tccaccttcc tccacccctc
     1561 agaaagtcga cccccggaag ctcacccgga acttgctcct cagcggagac aatgagctct
     1621 acccactcag cccagggaag gacatggagc ccaacggccc gtcgctgccc agggatgaag
     1681 ggcccccgac cccaagctct gccacgaagg tgccaccggc agagtacagg ctgtgcaacg
     1741 ggtcagacaa ggaatgtgtg tcccccaccg ccagggtcac caagaaggag actctcaagg
     1801 cgcagaagga gaactaccgg caggagaaga agcgcgccac acggcagctg ctcagcgctc
     1861 tgacagaccc cagcgtggtc atcatggctg acagcctgaa gggccccaaa ggtgagagcg
     1921 tgggctccat cacacagccc ctgcccagca gctacctgat cttcagggcc gcctccgagt
     1981 cagatggtcg ctgctggctg gacgccctgg agctggccct gcgctgctct agcctactga
     2041 gactgggcac ctgcaagccg ggccgagacg gggagccagg gacctcgcca gacgcatcac
     2101 cctcatcgct ctgtgggctg ccagcctcag ccaccgtcca cccagaccaa gacctgttcc
     2161 cactgaacgg gtcttccctg gagaacgatg cattctcaga caagtcggag agagagaacc
     2221 ctgaggagtc agataccgag acccaggacc atagccggaa gacggagagt ggcagcgacc
     2281 agtcagagac ccctggggcc ccggtgcgga gagggaccac ctatgtggag caggtccagg
     2341 aggagctggg ggagctgggc gaggcgtccc aggtggagac agtgtcagag gagaacaaga
     2401 gtctgatgtg gaccctgctg aagcagctac ggccaggcat ggacctgtcc cgcgtggtgc
     2461 tacccacgtt cgtactggag ccgcgctcct tcctgaacaa gctctccgac tactactacc
     2521 acgcagacct gctctccagg gctgcggtgg aggaggatgc ctacagccgc atgaagctgg
     2581 tgctgcggtg gtacctgtct ggcttctaca agaagcccaa gggaatcaag aagccgtaca
     2641 accccatcct gggggagacc ttccgctgct gctggttcca cccgcagact gacagccgca
     2701 cattctacat agcagagcag gtgtcccacc acccgcccgt gtctgccttc cacgtcagca
     2761 accggaagga cggcttctgc atcagtggca gcatcacagc caagtccagg ttttatggga
     2821 actcgctgtc ggcgctgctg gacggcaaag ccacgctcac cttcctgaac cgagccgagg
     2881 attacaccct taccatgccc tacgcccact gcaaaggaat cctgtatggc acgatgaccc
     2941 tggagctggg tgggaaggtc accatcgagt gtgcgaagaa caacttccag gcccagctgg
     3001 aattcaaact caagcccttc ttcgggggta gcaccagcat caaccagatc tcgggaaaga
     3061 tcacgtcggg agaggaagtc ctggcgagcc tcagtggcca ctgggacagg gacgtgttta
     3121 tcaaggagga agggagcgga agcagtgcgc ttttctggac cccgagcggg gaggtccgca
     3181 gacagaggct gaggcagcac acggtgccgc tggaggagca gacggagctg gagtccgaga
     3241 ggctctggca gcacgtcacc agggccatca gcaagggcga ccagcacagg gccacacagg
     3301 agaagtttgc actggaggag gcacagcggc agcgggcccg tgagcggcag gagagcctca
     3361 tgccctggaa gccgcagctg ttccacctgg accccatcac ccaggagtgg cactaccgat
     3421 acgaggacca cagcccctgg gaccccctga aggacatcgc ccagtttgag caagacggga
     3481 tcctgcggac cttgcagcag gaggccgtgg cccgccagac caccttcctg ggcagcccag
     3541 ggcccaggca cgagaggtct ggcccagacc agcggcttcg caaggccagc gaccagccct
     3601 ccggccacag ccaggccacg gagagcagcg gatccacgcc tgagtcctgc ccagagctct
     3661 cagacgagga gcaggatggt gactttgtcc ctggcggtga gagcccatgc cctcggtgca
     3721 ggaaggaggc gcggcggctg caggccctgc acgaggccat cctctccatc cgagaggccc
     3781 agcaggagct gcacaggcac ctctcggcca tgctgagctc cacggcacgg gcagcacagg
     3841 caccgacccc aggcctcctg cagagccccc gatcctggtt cctgctctgc gtgttcctgg
     3901 cgtgtcagct gttcattaac cacatcctca aactcgagga ctacaaagac catgacggtg
     3961 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     4021 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4081 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4141 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4201 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4261 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4321 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4381 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4441 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4501 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4561 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4621 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4681 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4741 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4801 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4861 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4921 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4981 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5041 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     5101 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5161 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5221 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5281 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     5341 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5401 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5461 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5521 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5581 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5641 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5701 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5761 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5821 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5881 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5941 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6001 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6061 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6121 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6181 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6241 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6301 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6361 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6421 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6481 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6541 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6601 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6661 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6721 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6781 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6841 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6901 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6961 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7021 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7501 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7741 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7861 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7921 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7981 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8041 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8161 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8221 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8281 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8581 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8941 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9001 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9061 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     9121 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9181 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     9241 aatctgctta gg
//
