LOCUS       sequence-203651-        5306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for red fluorescent Ca2+ indicator 
            K-GECO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5306)
  AUTHORS   Shen Y, Dana H, Abdelfattah AS, Patel R, Shea J, Molina RS, Rawal B,
            Rancic V, Chang YF, Wu L, Chen Y, Qian Y, Wiens MD, Hambleton N, 
            Ballanyi K, Hughes TE, Drobizhev M, Kim DS, Koyama M, Schreiter ER, 
            Campbell RE
  TITLE     A genetically encoded Ca(2+) indicator based on circularly 
            permutated sea anemone red fluorescent protein eqFP578.
  JOURNAL   BMC Biol. 2018 Jan 16;16(1):9. doi: 10.1186/s12915-018-0480-0.
  PUBMED    29338710
REFERENCE   2  (bases 1 to 5306)
  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: "BMC Biol.";
            date: "2018-01-16"; volume: "16(1):9. doi"; pages: " 
            10.1186/s12915-018-0480-0"
COMMENT     Sequence Label: pBAD-K-GECO1
FEATURES             Location/Qualifiers
     source          1..5306
                     /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"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1707..1724)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1877..1963
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2055..2082
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2101..2192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2193..2261,2262..3053)
                     /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(2411..2430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3224..3812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3713..3732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3966..3983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3998..4138
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4064..4083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4224..4246
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4402..5280)
                     /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 ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagaatgg gcagcgtgaa gctgatcccc agcctgacca ccgtgatcct cgtgaagtcc
      481 atgctgcgga agcggagctt cggcaacccc ttcaagtata atacggagac cctgtacccc
      541 gctgacggcg gcctggaagg cgcatgtgac atggccctga agctcgtggg cgggggccac
      601 ctgaactgca gccttgagac cacatacaga tccaagaaac ccgctacgaa cctcaagatg
      661 cccggtgtct acaacgtgga ccacagactg gaacgaatca aagaggccga cgatgagacc
      721 tacgtcgagc tgcacgaggt ggctgtggcc agatacgtgg gcctgggtgg tggcggaggt
      781 acaggcggga gtgtgagcga gctgattaag gagaacatgc caatgaagct gtacatggag
      841 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
      901 ggcacccaga ccatgagaat caaggtcgtc gagggcggcc ctctcccctt cgccttcgac
      961 atcctggcta ccagcttcat gtacggcagc agaaccttca tcaagcaccc tcctggcatc
     1021 cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
     1081 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
     1141 tacaacgtca aggttagagg gatgaacttc ccagccaacg gccctgtgat gcagaagaaa
     1201 acactcggct gggaggccag taatggccaa ctgactgaag agcagatcgc agaatttaaa
     1261 gaggctttct ccctatttga caaggacggg gatgggacga taacaaccaa ggagctgggg
     1321 acggtgatgc ggtctctggg gcagaacccc acagaagcgg agctgcagga catgatcaat
     1381 gaagtagatg ccgacggtga cggcacattc gacttccctg agttcctgac gatgatggca
     1441 agaaaaatga gttatagagt cactgaagag gaaattagag aagcgttccg cgtgtttgat
     1501 aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac agaccttgga
     1561 gagaagttaa cagatgagga ggttgatgaa atgatcaggg tagcagacat cgatggggat
     1621 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtagaa gcttggctgt
     1681 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1741 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1801 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1861 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1921 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1981 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2041 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2161 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2221 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2281 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2341 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2401 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2461 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2521 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2581 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2641 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2701 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2761 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2821 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2881 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2941 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3001 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3061 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3241 tgcgcgtaat ctgctgcttg caaacaaaac aaccaccgct accagcggtg gtttgtttgc
     3301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3361 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4021 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     4081 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4141 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4201 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4261 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     4321 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4381 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4441 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4501 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4561 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4621 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4681 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4741 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4801 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4861 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     4921 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4981 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     5041 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     5101 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     5161 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     5221 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     5281 acttttcata ctcccgccat tcagag
//
