LOCUS       V024091                 8755 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024091
VERSION     V024091
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..3501
                     /label="AGO1(NM_012199)"
                     /note="AGO1(NM_012199)"
                     /gene="AGO1"
     CDS             3517..3573
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3574..4284
                     /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"
     polyA_signal    4322..4546
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4592..5020
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5034..5363
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5430..6224
                     /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    6398..6519
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6568..6584)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6592..6608
                     /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(6616..6646)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6661..6682
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6970..7555)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7726..8586)
                     /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(8587..8691)
                     /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 atggaagcgg gaccctcggg agcagctgcg
      961 ggcgcttacc tgccccccct gcagcaggtg ttccaggcac ctcgccggcc tggcattggc
     1021 actgtgggga aaccaatcaa gctcctggcc aattactttg aggtggacat ccctaagatc
     1081 gacgtgtacc actacgaggt ggacatcaag ccggataagt gtccccgtag agtcaaccgg
     1141 gaagtggtgg aatacatggt ccagcatttc aagcctcaga tctttggtga tcgcaagcct
     1201 gtgtatgatg gaaagaagaa catttacact gtcacagcac tgcccattgg caacgaacgg
     1261 gtcgactttg aggtgacaat ccctggggaa gggaaggatc gaatctttaa ggtctccatc
     1321 aagtggctag ccattgtgag ctggcgaatg ctgcatgagg ccctggtcag cggccagatc
     1381 cctgttccct tggagtctgt gcaagccctg gatgtggcca tgaggcacct ggcatccatg
     1441 aggtacaccc ctgtgggccg ctccttcttc tcaccgcctg agggctacta ccacccgctg
     1501 gggggtgggc gcgaggtctg gttcggcttt caccagtctg tgcgccctgc catgtggaag
     1561 atgatgctca acattgatgt ctcagccact gccttttata aggcacagcc agtgattgag
     1621 ttcatgtgtg aggtgctgga catcaggaac atagatgagc agcccaagcc cctcacggac
     1681 tctcagcgcg ttcgcttcac caaggagatc aagggcctga aggtggaagt cacccactgt
     1741 ggacagatga agaggaagta ccgcgtgtgt aatgttaccc gtcgccctgc tagccatcag
     1801 acattcccct tacagctgga gagtggacag actgtggagt gcacagtggc acagtatttc
     1861 aagcagaaat ataaccttca gctcaagtat ccccatctgc cctgcctaca agttggccag
     1921 gaacaaaagc atacctacct tcccctagag gtctgtaaca ttgtggctgg gcagcgctgt
     1981 attaaaaagc tgaccgacaa ccagacctcg accatgataa aggccacagc tagatccgct
     2041 ccagacagac aggaggagat cagtcgcctg atgaagaatg ccagctacaa cttagatccc
     2101 tacatccagg aatttgggat caaagtgaag gatgacatga cggaggtgac agggcgagtg
     2161 ctgccggcgc ccatcttgca gtacggcggc cggaaccggg ccattgccac acccaatcag
     2221 ggtgtctggg acatgcgggg gaaacagttc tacaatggga ttgagatcaa agtctgggcc
     2281 atcgcctgct tcgcacccca aaaacagtgt cgagaagagg tgctcaagaa cttcacagac
     2341 cagctgcgga agatttccaa ggatgcgggg atgcctatcc agggtcaacc ttgtttctgc
     2401 aaatatgcac agggggcaga cagcgtggag cctatgttcc ggcatctcaa gaacacctac
     2461 tcagggctgc agctcattat tgtcatcctg ccagggaaga cgccggtgta tgctgaggtg
     2521 aaacgtgtcg gagatacact cttgggaatg gctacgcagt gtgtgcaggt gaagaacgtg
     2581 gtcaagacct cacctcagac tctgtccaac ctctgcctca agatcaatgt caaacttggt
     2641 ggcattaaca acatcctagt cccacaccag cgctctgccg tttttcaaca gccagtgata
     2701 ttcctgggag cagatgttac acacccccca gcaggggatg ggaaaaaacc ttctatcaca
     2761 gcagtggtag gcagtatgga tgcccacccc agccgatact gtgctactgt gcgggtacag
     2821 cgaccacggc aagagatcat tgaagacttg tcctacatgg tgcgtgagct cctcatccaa
     2881 ttctacaagt ccacccgttt caagcctacc cgcatcatct tctaccgaga tggggtgcct
     2941 gaaggccagc taccccagat actccactat gagctactgg ccattcgtga tgcctgcatc
     3001 aaactggaaa aggactacca gcctgggatc acttatattg tggtgcagaa acgccatcac
     3061 acccgccttt tctgtgctga caagaatgag cgaattggga agagtggtaa catcccagct
     3121 gggaccacag tggacaccaa catcacccac ccatttgagt ttgacttcta tctgtgcagc
     3181 cacgcaggca tccagggcac cagccgacca tcccattact atgttctttg ggatgacaac
     3241 cgtttcacag cagatgagct ccagatcctg acgtaccagc tgtgccacac ttacgtacga
     3301 tgcacacgct ctgtctctat cccagcacct gcctactatg cccgcctggt ggctttccgg
     3361 gcacgatacc acctggtgga caaggagcat gacagtggag aggggagcca catatcgggg
     3421 cagagcaatg ggcgggaccc ccaggccctg gccaaagccg tgcaggttca ccaggatact
     3481 ctgcgcacca tgtacttcgc tctcgaggga agcggagcta ctaacttcag cctgctgaag
     3541 caggctggag acgtggagga gaaccctgga cctatggtga gcaagggcga ggaggataac
     3601 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     3661 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     3721 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     3781 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     3841 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     3901 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     3961 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     4021 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     4081 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     4141 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     4201 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     4261 ggcatggacg agctgtacaa gtaatctaga gggcccgttt aaacccgctg atcagcctcg
     4321 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4381 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4441 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4501 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4561 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4621 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4681 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4741 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4921 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5041 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5101 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5161 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5221 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5281 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5341 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5401 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5461 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5521 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5581 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5641 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5701 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5761 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5821 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5881 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5941 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6001 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6061 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6121 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6181 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6241 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6301 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6361 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6541 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6721 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6781 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6841 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6901 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6961 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7021 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7081 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7141 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7201 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7261 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7321 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7381 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7441 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7501 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8701 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
