LOCUS       sequence-205943-        5485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A yellow fluorescent Ca2+ indicator variant with medium binding 
            affinity and kinetics for expression in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5485)
  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 5485)
  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-GECO2m
FEATURES             Location/Qualifiers
     source          1..5485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDARQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1521..1540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1774..1791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1806..1946
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1872..1891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2032..2054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2210..3088)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3115..3398
                     /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     3321..3340
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             3440..3451
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3606..3623
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3627..3659
                     /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             4721..4735
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(5000..5017)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5170..5256
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5348..5375
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5394..5485
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtgttgacg ccaggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1801 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     1861 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     1921 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     1981 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2041 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     2101 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     2161 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     2221 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     2281 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     2341 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     2401 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     2461 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     2521 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     2581 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     2641 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     2701 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     2761 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     2821 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     2881 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     2941 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     3001 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     3061 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     3121 ccaattgtcc atattgcatc agtcaggtcg tcactgcgtc ttttactggc tcttctcgct
     3181 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     3241 gacaaaatcg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     3301 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     3361 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg
     3421 aggaattaac catgggttta aagaggagaa aggtcatgaa caataacgat ctctttcagg
     3481 catcacgtcg gcgttttctg gcacaactcg gcagcttaac cgacgccggg acgctggggc
     3541 cgtcattgtt aacgccgcga cgtgcgactg cggcgcaagc ggcgactgac gcttctagag
     3601 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
     3661 gggatctgta cgacgaagat gataaggatc tcgccacaac ggccgactca tcacgtcgta
     3721 agtggaataa gacacacgca gtcagagcta taggccggct gagctcactc gagggttata
     3781 gttccgagaa aattgtgcct gttccaaagc aagggatatt aaaaggaaag gtcagcatgt
     3841 acctattgct gaaggatggc ggtcgttatc attgccagtt tgatacggta tataaagcta
     3901 aatcagtgcc ctccaagatg ccaggacggc attttataca gcataagctc cttagggagg
     3961 accgctctga tgctaaaaac cataaatggc aattaacaga acatgcaatc gcaggcatgg
     4021 acgagctgta caagggaggt acaggcggga gtatggtgag caagggcgag gggcaaccga
     4081 agcatgggct aaaggaggag atgacggtga agtaccacat ggaaggttgc gttaatggtc
     4141 acaaatttgt cattactgga gagggcattg gaaacccttt taagggtaag caaactgcaa
     4201 atttgtgtgt gatagaaggg ggcccgctgc cgttctcgga ggacatttta agtccgggtt
     4261 ttaaatatgg tgaccggatt ttcacagagt acccacagga tattgtagat tacttcaaga
     4321 actcatgtcc agagggctat acgtgggaaa ggagcttctt ctatgaggac ggagcggtct
     4381 gtcgatgcaa cgcggacata acagtctctg aaaaggagaa ctgcatctat cacaaaagta
     4441 tcttctatgg ggtgaatttt cccgccgacg gccccgtaat gaagaaaatg accactaatt
     4501 gggaagctga cacgcgtgac gacctgactg aagagcagat cgcagaattt aaagaggctt
     4561 tctccctatt tgacaaggac ggggatggga cgataacaac cgaggagctg gggacggtgt
     4621 tccggtctct ggggcagaac ccctcagaag cagagctgcg ggacatgatc aatgaagtag
     4681 atgccgacgg tgacggcaca atcggcttcc ctgagttcct gacgatgatg acaagaaaaa
     4741 tgaatgacac ggacagtgaa gaggaaatta gagaagcgtt ccgtgtgttt gataaggacg
     4801 gcaatggcta catcggcgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     4861 taacagatga agagtttgat gaaatgatca gggtagcaga ctccgacggg gatggtcagg
     4921 taaactacga agagtttgta caaatgatga cagcgaagta aaagcttggc tgttttggcg
     4981 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     5041 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     5101 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     5161 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5221 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5281 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     5341 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     5401 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5461 tcaataatat tgaaaaagga agagt
//
