LOCUS Exported 6706 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CaYin1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6706) AUTHORS Ding Y, Ai HW, Hoi H, Campbell RE TITLE Forster resonance energy transfer-based biosensors for multiparameter ratiometric imaging of Ca2+ dynamics and caspase-3 activity in single cells. JOURNAL Anal Chem. 2011 Dec 15;83(24):9687-93. doi: 10.1021/ac202595g. Epub 2011 Nov 28. PUBMED 22080726 REFERENCE 2 (bases 1 to 6706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6706 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..284 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 208..227 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 311..333 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 353..370 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 374..406 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 419..439 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(2387..2408) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3107..3124) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 3277..3363 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3455..3482 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3501..3592 /gene="bla" /label=AmpR promoter CDS 3593..4453 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3811..3830) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4624..5212 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5113..5132 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5366..5383 /label=L4440 /note="L4440 vector, forward primer" misc_feature 5398..5538 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(5464..5483) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5624..5646 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(5802..6680) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct 61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca 121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg 181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg 241 atcctacctg acgcttttta tcgcactctc tactgtttct ccatacccgt ttttttgggc 301 tagaaataat tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca 361 tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga 421 gaacctgtac ttccagggct cgagcatggt gagcaagggc gaggaggtca tcaaagagtt 481 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg 541 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg 601 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc 661 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt 721 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc 781 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc 841 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta 901 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg 961 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg 1021 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt 1081 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag 1141 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa 1201 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat 1261 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac 1321 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc 1381 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga 1441 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca 1501 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt 1561 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg 1621 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga 1681 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact 1741 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac 1801 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgc gcatgcatct 1861 gacagaagag cagattgcag agttcaaaga agccttctca ttattggaca aggatgggga 1921 cggcaccatc accacaaagg aacttggcac cgctctgagg tcgcttggac aaaacccaac 1981 ggaagcagaa ttgcaggata tgatcaatga agtcgatgct gatggcaatg gaacgattta 2041 ctttcctgaa tttcttacta tgatggctag aaaaatgaag gacacagaca gcgaagagga 2101 aatccgagaa gcattccgtg tttttgacaa ggatgggaac ggctacatca gcgctgctga 2161 attacgtcac gtcatgacaa acctcgggga gaagttaaca gatgaagaag ttgatgaaat 2221 gataagggaa gcagatatcg atggtgatgg ccaagtaaac tatgaagagt ttgtacaaat 2281 gatgacagca aaggggggga agaggcgctg gcagaaaaca ggccatgctg tccgagcatt 2341 cggaagactg aagaagatcg agctcttcac cggggtggtg cccatcctgg tcgagctgga 2401 cggcgacgta aacggccaca ggttcagcgt gcgcggcgag ggcgagggcg atgccaccaa 2461 cggcaagctg atcctgaagt tcatctgcac ctccggcgag ctgcccgtgc cctggcccac 2521 cctcgtgacc accctgtctt acggcgtgca gtgcttcgcc cgctaccccg accacatgaa 2581 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 2641 cttcaaggac gacggcagct acaggacccg cgccgaggtg aagttcgagg gcgacaccct 2701 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 2761 caagctggag tacaacatga acgcgtggga cgcgtatatc acggccgaca agcagaagaa 2821 cggcatcaaa gcgaacttca agatcgagca caacgtcaag gacggcggcg tgcagctcgc 2881 cgacgcgtac cagcagaaca cccccatcgg cggcggcccc gtgctgctgc ctgacaacca 2941 ctacctgagc ttccagagca agctgttcaa agacccctac gagcagcgcg atcacatggt 3001 cctgctggag ttcgttaccg ccgccgggat aactcccggc atggacgagc tgtacaagta 3061 agaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta 3121 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg 3181 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg 3241 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg 3301 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca 3361 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga 3421 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt 3481 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct 3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 3661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 3721 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 3781 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga 3841 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 3901 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 3961 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 4021 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 4081 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 4141 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 4201 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 4261 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 4321 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 4381 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 4441 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 4501 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4561 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4621 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 4681 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 4741 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 4801 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4861 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4921 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4981 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5041 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5101 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5161 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5221 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 5281 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 5341 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct 5401 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct 5461 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt 5521 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 5581 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 5641 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag 5701 gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta 5761 tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct 5821 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat 5881 tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg 5941 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg 6001 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc 6061 gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg 6121 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta 6181 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa 6241 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc 6301 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc 6361 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc 6421 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt 6481 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg 6541 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc 6601 tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga 6661 tcattttgcg cttcagccat acttttcata ctcccgccat tcagag //