LOCUS Exported 5471 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTorPE-GEM-GECO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5471) AUTHORS Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, Abdelfattah AS, Fujiwara M, Ishihara T, Nagai T, Campbell RE TITLE An Expanded Palette of Genetically Encoded Ca2+ Indicators. JOURNAL Science. 2011 Sep 8. PUBMED 21903779 REFERENCE 2 (bases 1 to 5471) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5471 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..285 /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 327..338 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 493..510 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 514..546 /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 550..573 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 601..657 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 870..891 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1255..1274) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind complement(1872..1889) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2042..2128 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2220..2247 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2266..2357 /gene="bla" /label=AmpR promoter CDS 2358..3218 /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(2576..2595) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3389..3977 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3878..3897 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4131..4148 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4163..4303 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4229..4248) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4389..4411 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4567..5445) /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 tcgcaactct ctactgtttc tccatacccg ttttttgggc 301 taacaggagg aattaaccat gggtttaaag aggagaaagg tcatgaacaa taacgatctc 361 tttcaggcat cacgtcggcg ttttctggca caactcggca gcttaaccga cgccgggacg 421 ctggggccgt cattgttaac gccgcgacgt gcgactgcgg cgcaagcggc gactgacgct 481 tctagaggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa 541 atgggtcggg atctgtacga cgatgatgat aaggatctcg ccacaatggt cgactcatca 601 cgtcgtaagt ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcacca 661 gagaacgtgt atataaaggc cgacgagcag aagaacggca tcaaggcgta cttcaagatc 721 cgccacaaca tcgagggcgg cggcgtgcag ctcgcctacc actaccagca gatcaccccc 781 ataggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccatactt 841 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 901 gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat ggtgagcaag 961 ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg agctggacgg cgacgtaaac 1021 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc 1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 1141 ctgtcctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 1201 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac 1261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 1321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 1381 agcacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta 1441 tttgacaagg acggggatgg gacgataaca accaaggagc tggggacggt gatgcggtct 1501 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac 1561 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcacctaa aatgcaggac 1621 acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga cggcaatggc 1681 tacatcggcg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat 1741 gaagaggttg atgaaatgat cagggtagca gacatcgatg gnganggtca ggtaaactac 1801 gaagagtttg tacaaatgat gacagcgaag tagaagcttg gctgttttgg cggatgagag 1861 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat 1921 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa 1981 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca 2041 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc 2101 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca 2161 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca 2221 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct 2281 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 2341 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 2401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 2461 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 2521 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 2581 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact 2641 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 2701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 2761 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 2821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 2881 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 2941 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 3001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 3061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 3121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 3181 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 3241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 3301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 3361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 3421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 3481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 3541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 3601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 3661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 3721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 3781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 3841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 3901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 3961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 4021 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 4081 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 4141 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 4201 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 4261 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac 4321 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 4381 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 4441 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga 4501 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt 4561 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa 4621 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc 4681 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt 4741 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg 4801 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc 4861 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc 4921 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag 4981 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg 5041 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt 5101 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac 5161 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg 5221 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac 5281 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat 5341 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt 5401 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc 5461 gccattcaga g //