LOCUS       sequence-37238-e        5477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5477)
  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 30;333(6051):1888-91. doi: 10.1126/science.1208592
  PUBMED    21903779
REFERENCE   2  (bases 1 to 5477)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2011-09-30- 30"; volume: "333"; 
            issue: "6051"; pages: "1888-91"
COMMENT     Sequence Label: pTorPE-R-GECO1
FEATURES             Location/Qualifiers
     source          1..5477
                     /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             join(550..558,559..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="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1353..1372
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1396..1839
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1878..1895)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2048..2134
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2226..2253
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2272..2363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2364..2432,2433..3224)
                     /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(2582..2601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3395..3983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3884..3903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4137..4154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4169..4309
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4235..4254)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4395..4417
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4573..5451)
                     /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 aaaaacgcga 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 cgactcttca
      601 cgtcgtaagt ggaataaggc aggtcacgca gtcagagcta taggtcggct gagctcaccc
      661 gtggtttccg agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagggg
      721 ctgaggctga aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag
      781 aagcccgtgc agctgcccgg cgcctacatc gtcgacatca agttggacat cgtgtcccac
      841 aacgaggact acaccatcgt ggaacagtgc gaacgcgccg agggccgcca ctccaccggc
      901 ggcatggacg agctgtacaa gggaggtaca ggcgggagtc tggtgagcaa gggcgaggag
      961 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1021 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga ggcctttcag
     1081 accgctaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1141 cctcagttca tgtacggctc caaggcctac attaagcacc cagccgacat ccccgactac
     1201 ttcaagctgt ccttccccga gggcttcagg tgggagcgcg tgatgaactt cgaggacggc
     1261 ggcattattc acgttaacca ggactcctcc ctgcaggacg gcgtattcat ctacaaggtg
     1321 aagctgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     1381 tgggaggcta cgcgtgacca actgactgaa gagcagatcg cagaatttaa agaggctttc
     1441 tccctatttg acaaggacgg ggatgggacg ataacaacca aggagctggg gacggtgatg
     1501 cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa tgaagtagat
     1561 gccgacggtg acggcacatt cgacttccct gagttcctga cgatgatggc aagaaaaatg
     1621 aatgacacag acagtgaaga ggaaattaga gaagcgttcc gcgtgtttga taaggacggc
     1681 aatggctaca tcggcgcagc agagcttcgc cacgtgatga cagaccttgg agagaagtta
     1741 acagatgagg aggttgatga aatgatcagg gtagcagaca tcgatgggga tggtcaggta
     1801 aactacgaag agtttgtaca aatgatgaca gcgaagtaga agcttggctg ttttggcgga
     1861 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1921 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     1981 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     2041 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2101 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     2161 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     2221 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     2281 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2341 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2401 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2461 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2521 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2581 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2641 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2701 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2761 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2821 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2881 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2941 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3001 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3061 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3121 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3181 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4081 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4141 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4201 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4261 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     4321 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4381 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4441 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
     4501 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     4561 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4621 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4681 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4741 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4801 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     4861 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     4921 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     4981 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     5041 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     5101 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     5161 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     5221 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     5281 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     5341 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     5401 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     5461 actcccgcca ttcagag
//
