LOCUS       sequence-37236-e        5474 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 5474)
  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 5474)
  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-GEX-GECO1
FEATURES             Location/Qualifiers
     source          1..5474
                     /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="RRKWNKTGHAVRAIGRLSS"
     primer_bind     870..891
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1258..1277)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1393..1836
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAELKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDLPEFQTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1875..1892)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2045..2131
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2223..2250
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2269..2360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2361..2429,2430..3221)
                     /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(2579..2598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3392..3980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3881..3900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4134..4151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4166..4306
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4232..4251)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4392..4414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4570..5448)
                     /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 gagctcactc
      661 gagaacgtct atatcaaggc cgacgagcag aagaacggca tcaaggcgaa cttcaagatc
      721 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
      781 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccatactt
      841 tcgaaagacc ccaacgagaa gcgtgatcac atggtcctgc tggagttcgt gaccgccgcc
      901 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc
      961 aagggcgagg agctgttcac cggggtggtg cccatccagg tcgagctgga cggcgacgta
     1021 aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg
     1081 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1141 accctgtcct atggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1201 ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac
     1261 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1321 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1381 tacaacacgc gtgaccaact gactgaagag cagatcgcag aacttaaaga ggctttctcc
     1441 ctatttgaca aggacgggga tgggacgata acaaccaagg agctggggac ggtgatgcgg
     1501 tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc
     1561 gacggtgacg gcacaatcga cctccctgag ttccagacaa tgatggcaag aaaaatgaat
     1621 gacacagaca gtgaagagga aattagagaa gcgttccgtg tgtttgataa ggacggcaat
     1681 ggctacatcg gcgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca
     1741 gatgaagagg tagatgaaat gatcagggta gcagacatcg atggggatgg tcaggtaaac
     1801 tacgaagagt ttgtacaaat gatgacagcg aagtagaagc ttggctgttt tggcggatga
     1861 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     1921 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     1981 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     2041 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2101 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     2161 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     2221 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     2281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2581 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     2641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4141 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4201 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4261 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4321 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4381 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4441 gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat
     4501 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     4561 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     4621 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     4681 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     4741 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     4801 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     4861 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     4921 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     4981 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     5041 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     5101 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     5161 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     5221 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     5281 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     5341 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     5401 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     5461 cccgccattc agag
//
