LOCUS       sequence-37233-e        5471 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 5471)
  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 5471)
  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-GEM-GECO1
FEATURES             Location/Qualifiers
     source          1..5471
                     /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(1255..1274)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1872..1889)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2042..2128
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2220..2247
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2266..2357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2358..2426,2427..3218)
                     /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(2576..2595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3389..3977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3878..3897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4131..4148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4163..4303
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4229..4248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4389..4411
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4567..5445)
                     /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 gagctcacca
      661 gagaacgtgt atataaaggc cgacgagcag aagaacggca tcaaggcgta cttcaagatc
      721 cgccacaaca tcgagggcgg cggcgtgcag ctcgcctacc actaccagca gatcaccccc
      781 ataggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccatactt
      841 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      901 gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat ggtgagcaag
      961 ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg agctggacgg cgacgtaaac
     1021 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1141 ctgtcctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1201 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     1261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1381 agcacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta
     1441 tttgacaagg acggggatgg gacgataaca accaaggagc tggggacggt gatgcggtct
     1501 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     1561 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcacctaa aatgcaggac
     1621 acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga cggcaatggc
     1681 tacatcggcg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     1741 gaagaggttg atgaaatgat cagggtagca gacatcgatg gnganggtca ggtaaactac
     1801 gaagagtttg tacaaatgat gacagcgaag tagaagcttg gctgttttgg cggatgagag
     1861 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     1921 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     1981 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2041 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2101 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     2161 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     2221 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     2281 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2341 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2461 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2521 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2581 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2641 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2761 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2881 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2941 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3181 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4021 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4081 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4141 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4201 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4261 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     4321 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4381 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4441 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga
     4501 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     4561 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     4621 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     4681 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     4741 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     4801 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     4861 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     4921 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     4981 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     5041 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     5101 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     5161 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     5221 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     5281 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     5341 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     5401 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
     5461 gccattcaga g
//
