LOCUS       sequence-37246-e        5480 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 5480)
  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 5480)
  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-G-GECO1.2
FEATURES             Location/Qualifiers
     source          1..5480
                     /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"
     CDS             1399..1842
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1881..1898)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2051..2137
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2229..2256
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2275..2366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2367..2435,2436..3227)
                     /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(2585..2604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3398..3986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3887..3906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4140..4157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4172..4312
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4238..4257)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4398..4420
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4576..5454)
                     /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 tcaaggcgta cttcaagatc
      721 cgccacaaca tcgagggcgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
      781 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccatgctt
      841 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      901 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag cgaatccatg
      961 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tccaggtcga gctggacggc
     1021 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc
     1081 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1141 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1201 cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc
     1261 aagggcgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1321 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1381 ctggagtaca acacgcgtga ccaactgact gaagagcaga tcgcagaatt taaagaggct
     1441 ttctccctat ttgacaagga cggggatggg acgataacaa ccaaggagct ggggacggtg
     1501 atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat caatgaagta
     1561 gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa
     1621 atgaaagaca cagacagtga agaagaaatt agagaagcgt tccgtgtgtt tgataaggac
     1681 ggcaatggct acatcggcgc agcagagctt cgccacgtga tgacaaacct tggagagaag
     1741 ttaacagatg aagaggttga tgaaatgatc agggtagcag acatcgatgg ggatggtcag
     1801 gtaaactacg aagagtttgt acaaatgatg acagcgaagt agaagcttgg ctgttttggc
     1861 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     1921 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     1981 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     2041 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2101 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     2161 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     2221 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt
     2281 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2341 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2401 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2461 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2521 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2581 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     2641 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2701 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2761 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2821 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2881 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2941 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3001 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3061 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3121 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3181 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3241 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3301 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3361 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3421 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3481 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3541 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3601 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3661 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3721 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3781 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3841 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3901 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3961 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4021 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4081 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4141 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4201 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4261 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4321 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4381 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4441 acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat aatgtgcctg
     4501 tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt
     4561 ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     4621 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     4681 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     4741 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     4801 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     4861 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     4921 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     4981 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     5041 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     5101 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     5161 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     5221 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     5281 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     5341 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     5401 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     5461 catactcccg ccattcagag
//
