LOCUS Exported 9929 bp ds-DNA circular SYN 21-APR-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pHRSIN-CSW-AarI_AD aka bCW4_83_AarI pHR_primers10_11 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9929) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 21, 2016 from SnapGene 3.0.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9929 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 96..7517 /label=match to pHR-CMVLacZ /note="match to pHR-CMVLacZ" primer_bind complement(320..337) /label=SP6 /note="SP6" CDS 1231..2091 /codon_start=1 /label=bla (AmpR) /note="bla" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" CDS 1429..2088 /label=AmpR /note="AmpR" rep_origin 2186..2868 /label=ColE1 origin /note="ColE1 origin" promoter 3084..3410 /label=SV40 - Invitrogen /note="SV40 - Invitrogen promoter" promoter 3229..3425 /label=SV40 early promoter /note="SV40 early promoter" polyA_site 5105..5123 /label=SV40 pA /note="SV40 pA" polyA_signal complement(5138..5329) /label=SV40 late polyA /note="SV40 late polyA" polyA_site 5199..5330 /label=SV40 early poly A /note="SV40 early poly A" polyA_site 5199..5329 /label=SV40 early poly A (2) /note="SV40 early poly A" repeat_region 7518..7697 /label=cPPT /note="cPPT" primer_bind 7577..7598 /note="QB2741" primer_bind 7673..7696 /note="cPPT_5'seq" promoter 7730..8223 /label=SFFV /note="SFFV" primer_bind 8143..8162 /label=cw10 /note="cw10" misc_feature 8262..8265 /label=Part A /note="Part A" misc_feature complement(8270..8276) /label=AarI(1) /note="AarI" misc_feature 8288..8296 /note="Kozak" CDS 8297..9229 /codon_start=1 /translation="METDTLLLWVLLLWVPGSTGDMVSKGEELFTGVVPILVELDGDVN GHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHD FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL EYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKNAVGQDTQEVIVVPHSLPFKV VVISAILALVVLTIISLIILIMLWQKKPR" misc_feature 8297..8359 /note="signal sequence" misc_feature 8360..9076 /note="EGFP" misc_feature 9143..9211 /note="transmembrane" misc_feature 9256..9262 /label=AarI /note="AarI" misc_feature 9267..9270 /label=Part D /note="Part D" misc_feature 9302..9309 /label=SbfI /note="SbfI" enhancer 9338..9929 /label=WPRE /note="WPRE" misc_feature complement(9361..9384) /label=cw11 /note="cw11" primer_bind 9392..9412 /label=QB0112 WPRE-R binding site /note="QB0112 WPRE-R binding site" ORIGIN 1 cgataccgtc gaccaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 61 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc 121 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 181 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 241 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct 301 agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt 361 tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc 421 tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt 481 tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc 541 aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac 601 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg 661 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt 721 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct 781 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg 841 tcgaatggtg cactctcagt acaatctagc tctgatgccg catagttaag ccagccccga 901 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 961 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 1021 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 1081 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 1141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 1201 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 1261 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 1321 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 1381 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 1441 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 1501 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 1561 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 1621 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 1681 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 1741 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 1801 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 1861 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 1921 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 1981 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 2041 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 2101 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 2161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 2401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 3001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 3061 gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt 3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3421 gcaaaaagct tggacacaag acaggcttgc gagatatgtt tgagaatacc actttatccc 3481 gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat gtgaaatact ggtttttagt 3541 gcgccagatc tctataatct cgcgcaacct attttcccct cgaacacttt ttaagccgta 3601 gataaacagg ctgggacact tcacatgagc gaaaaataca tcgtcacctg ggacatgttg 3661 cagatccatg cacgtaaact cgcaagccga ctgatgcctt ctgaacaatg gaaaggcatt 3721 attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact ggcgcgtgaa ctgggtattc 3781 gtcatgtcga taccgtttgt atttccagct acgatcacga caaccagcgc gagcttaaag 3841 tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt tattgatgac ctggtggata 3901 ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa agcgcacttt gtcaccatct 3961 tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt tgttgatatc ccgcaagata 4021 cctggattga acagccgtgg gatatgggcg tcgtattcgt cccgccaatc tccggtcgct 4081 aatcttttca acgcctggca ctgccgggcg ttgttctttt taacttcagg cgggttacaa 4141 tagtttccag taagtattct ggaggctgca tccatgacac aggcaaacct gagcgaaacc 4201 ctgttcaaac cccgctttaa acatcctgaa acctcgacgc tagtccgccg ctttaatcac 4261 ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa ccatccctca ctggtatcgc 4321 atgattaacc gtctgatgtg gatctggcgc ggcattgacc cacgcgaaat cctcgacgtc 4381 caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg atgatttata cgatacggtg 4441 attggctacc gtggcggcaa ctggatttat gagtgggccc cggatctttg tgaaggaacc 4501 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt 4561 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt 4621 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga 4681 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca 4741 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga 4801 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat 4861 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt 4921 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca 4981 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt 5041 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa 5101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 5161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga 5341 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca 5401 attacctagt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 5461 agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc 5521 ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga 5581 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta 5641 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag 5701 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg 5761 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt 5821 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag 5881 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc 5941 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 6001 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 6061 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 6121 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga 6181 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg 6241 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga 6301 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc 6361 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg 6421 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca 6481 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc 6541 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa 6601 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gtgatcttca 6661 gacctggacg atatatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt 6721 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga 6781 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac 6841 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt 6901 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac 6961 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga 7021 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc 7081 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat 7141 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc 7201 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt 7261 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt 7321 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag 7381 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag 7441 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt 7501 gaacggatct cgacggtcgc caaatggcag tattcatcca caattttaaa agaaaagggg 7561 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 7621 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 7681 gcagagatcc agtttggatc gataagcttg atatcgaatt cctgcagccc cgataaaata 7741 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggttatg 7801 gcaagctagc tgcagtaacg ccattattgc aaggcatgga aaaataccaa accaagaata 7861 gagaagttca gatcaagggc gggtacatga aaatagctaa cgtagggcca aacaggatat 7921 ctgcggtgag cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg 7981 gccccggccc gaggccaaga gcagatggtc cccagatatg gcccaaccct cagcagtttc 8041 ttaagaccca tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat 8101 ttgaattaac caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc 8161 tataaaagag ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc 8221 gggggggatc tggagctctc gagaattctc acgcgtcaag tggagcaagg caggtggaca 8281 gtggatcgcc gccaccatgg agaccgacac cctcttgctc tgggtcctgc tgctgtgggt 8341 tccgggtagt accggagaca tggtttctaa gggtgaagaa ttgttcaccg gagttgtgcc 8401 catcctggtt gagctggacg gagacgttaa cggacacaag ttctcagtga gcggcgaagg 8461 agagggcgac gctacgtacg gtaaactgac gttgaagttc atctgtacaa caggaaagct 8521 gccagtgccc tggcctacac tcgtgaccac cctcacctac ggagtccagt gcttctcccg 8581 gtaccccgac cacatgaaac agcacgattt cttcaagtcc gcgatgcctg agggctatgt 8641 tcaggaacgc accatattct tcaaggacga cggtaactat aaaaccaggg ctgaagttaa 8701 gttcgagggg gacacgttgg taaatagaat tgagttgaaa ggaatcgatt tcaaagaaga 8761 cggcaatatt ttgggccaca agctggagta caactacaac agccacaacg tgtatattat 8821 ggctgataag cagaagaacg gcatcaaggt gaacttcaaa atccgacaca atatcgagga 8881 tggctctgtg caactggccg accattacca gcaaaacacg cctatcggcg acggtccggt 8941 tttgctgcca gataaccact acctctccac gcaatcagcc ctctccaagg atcctaacga 9001 aaaaagagac catatggttc tgcttgaatt cgtgacggct gctgggatca ccctgggaat 9061 ggatgagctg tataagaacg ccgtgggcca ggatacgcag gaggtgatcg tcgtgcccca 9121 cagcttgccc ttcaaggtcg tggttattag tgctatcctg gcgttggtgg tgctgactat 9181 tatctccctt atcatactta tcatgctgtg gcaaaaaaaa ccccgataag atccttgact 9241 tgcggccgca actcccacct gcaacatgcg tgactgactg aggccgcgac tctagagtcg 9301 acctgcaggc atgcaagctt gatatcaagc ttatcgataa tcaacctctg gattacaaaa 9361 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 9421 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 9481 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 9541 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 9601 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 9661 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 9721 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 9781 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 9841 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 9901 ggatctccct ttgggccgcc tccccgcat //