LOCUS       sequence-199209-        5486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A yellow fluorescent Ca2+ indicator variant with low binding 
            affinity and fast kinetics for expression in bacterial .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5486)
  AUTHORS   Zhao Y, Bushey D, Zhao Y, Schreiter ER, Harrison DJ, Wong AM, 
            Campbell RE
  TITLE     Inverse-response Ca(2+) indicators for optogenetic visualization of 
            neuronal inhibition.
  JOURNAL   Sci Rep. 2018 Aug 6;8(1):11758. doi: 10.1038/s41598-018-30080-x.
  PUBMED    30082904
REFERENCE   2  (bases 1 to 5486)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2018-08-6"; volume: "8(1):11758. doi"; pages: " 
            10.1038/s41598-018-30080-x"
COMMENT     Sequence Label: pTorPE-Y-GECO2f
FEATURES             Location/Qualifiers
     source          1..5486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1608..1622
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(1887..1904)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2057..2143
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2235..2262
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2281..2372
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2373..2441,2442..3233)
                     /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(2591..2610)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3404..3992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3893..3912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4146..4163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4178..4318
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4244..4263)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4404..4426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4582..5460)
                     /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 cgaagatgat aaggatctcg ccacaacggc cgactcatca
      601 cgtcgtaagt ggaataagac acacgcagtc agagctatag gccggctgag ctcactcgag
      661 ggttatagtt ccgagaaaat tgtgcctgtt ccaaagcaag ggatattaaa aggaaaggtc
      721 agcatgtacc tattgctgaa ggatggcggt cgttatcatt gccagtttga tacggtatat
      781 aaagctaaat cagtgccctc caagatgcca ggacggcatt ttatacagca taagctcctt
      841 agggaggacc gctctgatgc taaaaaccat aaatggcaat taacagaaca tgcaatcgca
      901 ggcatggacg agctgtacaa gggaggtaca ggcgggagta tggtgagcaa gggcgagggg
      961 caaccgaagc atgggctaaa ggaggagatg acggtgaagt accacatgga aggttgcgtt
     1021 aatggtcaca aatttgtcat tactggagag ggcattggaa acccttttaa gggtaagcaa
     1081 actgcaaatt tgtgtgtgat agaaggaggc ccgctgccgt tctcggagga cattttaagt
     1141 ccgggtttta aatatggtga ccggattttc acagagtacc cacaggatat tgtagattac
     1201 ttcaagaact catgtccaga gggctatacg tgggaaagga gcttcttcta tgaggacgga
     1261 gcggtctgtc gatgcaacgc ggacataaca gtctctgaaa aggagaactg catctatcac
     1321 aaaagtatct tctatggggt gaattttccc gccgacggcc ccgtaatgaa gaaaatgacc
     1381 actaattggg aagctgacac gcgtgaccaa ctgactgaag agcagatcgc agaatttaaa
     1441 gaggctttct ccctatttga caaggacggg gatgggacga tgacaaccga ggagctgggg
     1501 acggtgctgc ggtctctggg gcagaacccc tcagaagcag agctgcggga catgatcaat
     1561 gaagtagatg ccgacggtga cggcacaatc ggcttccctg agttcctgac gatgatgaca
     1621 agaaaaatga atgacacgga cagtgaagag gaaattagag aagcgttccg tgtgtttgat
     1681 aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac aaaccttgga
     1741 gagaagttaa cagatgaaga gtttgatgaa atgatcaggg tagcagactc cgacggggat
     1801 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag caaagtgaaa gcttggctgt
     1861 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1921 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1981 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2041 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2101 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2161 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2221 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     2281 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2341 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2401 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2461 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2521 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2581 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2641 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2701 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2761 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2821 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2881 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2941 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3001 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3061 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3121 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3181 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3241 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3301 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3541 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4141 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4201 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     4261 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4441 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     4501 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4561 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4621 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4681 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4741 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4801 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4861 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4921 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4981 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     5041 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     5101 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     5161 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     5221 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     5281 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     5341 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     5401 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     5461 acttttcata ctcccgccat tcagag
//
