LOCUS Exported 7496 bp ds-DNA circular SYN 02-NOV-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7496) AUTHORS Nordine Helassa TITLE Direct Submission JOURNAL Exported Nov 2, 2018 from SnapGene 4.2.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7496 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..467 /direction=RIGHT /label=M13 origin rep_origin complement(17..457) /direction=LEFT /label=F1 ori CDS complement(563..1375) /label=KanR rep_origin 1475..2103 /direction=RIGHT /label=ColE1 origin CDS complement(3281..4237) /label=LacI primer_bind 4751..4770 /label=T7 misc_binding 4770..4792 /label=LacO CDS 4839..5492 /label=GST CDS 5547..5564 /label=Thrombin site CDS 5580..5624 /label=S-tag CDS 5694..7256 /codon_start=1 /label=iGluSnFR /translation="AAGSTLDKIAKNGVIVVGHKESSVPFSYYDNQQKVVGYSQDYSNA IVEAVKKKLNKPDLQVKLIPITSQNRIPLLQNGTFDFECGSTTNNVERQKQAAFSDTIF VVGTRLLTKKGGDIKDFANLKDKAVVVTSGTTSEVLLNKLNEEQKMNMRIISAKDHGDS FRTLESGRAVAFMMDDVLLAGERAKAKKPDNWEIVGKPQSQEAYGCMLRKDDPQFKKLM DDTIAQVQTSGEAEKWFDKWFKNPILVSHNVYIMADKQRNGIKANFKIRHNIEDGGVQL AYHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELY KGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTG KLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRA EVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNNPLNMNFELSDEMKALFKEPNDK ALK" primer_bind 5730..5774 /label=Mutagenic Primer primer_bind 6653..6674 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(6780..6801) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7041..7060) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 7324..7341 /label=6His CDS 7327..7344 /label=6His CDS 7330..7347 /label=6His ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3181 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3241 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3361 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3421 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3481 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3541 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3601 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3661 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3721 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3781 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3841 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3901 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3961 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4021 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4081 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4141 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4201 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4261 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4321 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4381 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4441 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4501 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4561 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4621 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4681 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4741 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4801 gaaataattt tgtttaactt taagaaggag atatacatat gtcccctata ctaggttatt 4861 ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat 4921 atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat 4981 tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt 5041 ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag 5101 agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga 5161 gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg 5221 aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg 5281 taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa 5341 tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac 5401 aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag 5461 ccacgtttgg tggtggcgac catcctccaa aatcggatgg ttcaactagt ggttctggtc 5521 atcaccatca ccatcactcc gcgggtctgg tgccacgcgg tagtactgca attggtatga 5581 aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcccagat ctgggtaccg 5641 gtggtggctc cggtgatgac gacgacaaga gtcccatggg atatcgggga tccgccgcag 5701 gcagcacgct ggacaaaatc gccaaaaacg gtgtgattgt cgtcggtcac aaggaatctt 5761 cagtgccttt ctcttattac gacaatcagc aaaaagtggt gggttactcg caggattact 5821 ccaacgccat tgttgaagca gtgaaaaaga aactcaacaa accggacttg caggtaaaac 5881 tgattccgat tacctcacaa aaccgtattc cactgctgca aaacggcact ttcgattttg 5941 aatgtggttc taccaccaac aacgtcgaac gccaaaaaca ggcggctttc tctgacacta 6001 ttttcgtggt cggtacgcgc ctgttgacca aaaagggtgg cgatatcaaa gattttgcca 6061 acctgaaaga caaagccgta gtcgtcactt ccggcactac ctctgaagtt ttgctcaaca 6121 aactgaatga agagcaaaaa atgaatatgc gcatcatcag cgccaaagat cacggtgact 6181 ctttccgcac cctggaaagc ggtcgtgccg ttgcctttat gatggatgac gtgctgctgg 6241 ccggtgaacg tgcgaaagcg aagaaaccag acaactggga aatcgtcggc aagccgcagt 6301 ctcaggaggc ctacggttgt atgttgcgta aagatgatcc gcagttcaaa aagctgatgg 6361 atgacaccat cgctcaggtg cagacctccg gtgaagcgga aaaatggttt gataagtggt 6421 tcaaaaatcc aattctggtg agccacaacg tctatatcat ggccgacaag cagaggaacg 6481 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgcct 6541 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 6601 acctgagcac ccagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc 6661 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg 6721 gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc 6781 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg 6841 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg 6901 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc 6961 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg 7021 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg 7081 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca 7141 acatcctggg gcataagctg gagtacaact ttaacaaccc cctgaacatg aatttcgaac 7201 tgtcagacga aatgaaagca ctgttcaaag aaccgaatga caaggcactg aaataagaat 7261 tctgtacagg ccttggcgcg cctgcaggcg agctccgtcg acaagcttgc ggccgcactc 7321 gagcaccacc accaccacca ccaccactaa ttgattaata cctaggctgc taaacaaagc 7381 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg 7441 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat //