LOCUS       Exported                5515 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTorPE-G-GECO1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5515)
  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 8.
  PUBMED    21903779
REFERENCE   2  (bases 1 to 5515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5515
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             511..543
                     /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             547..570
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             598..654
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     867..888
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1255..1274)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1390..1830
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFSEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(1916..1933)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2086..2172
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2264..2291
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2310..2401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2402..3262
                     /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(2620..2639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3433..4021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3922..3941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4175..4192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4207..4347
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4273..4292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4433..4455
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4611..5489)
                     /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 tttactggca caactcggca acttaaccga cgccgggacg
      421 ctggggccgt cattgttaac gccgcgacgt gcaactgcgg cgcaagcggc gactgacgct
      481 tctagaggtt ctcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg
      541 ggtcgggatc tgtacgacga tgatgataag gatctcgcca caatggtcga ctcatcacgt
      601 cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag
      661 aacgtctata tcaaggccga cgagcagaag aacggcatca aggcgtactt caagatccgc
      721 cacaacatcg agggcggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
      781 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc catactttcg
      841 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      901 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
      961 ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg agctggacgg cgacgtaaac
     1021 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1141 ctgacctacg 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 aacacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta
     1441 tttgacaagg acggggatgg gacgataaca accaaggagc tggggacggt gatgcggtct
     1501 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     1561 ggtgacggca caatcgactt ctctgagttc ctgacaatga tggcaagaaa aatgaaagac
     1621 acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga cggcaatggc
     1681 tacatcggcg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     1741 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     1801 gaagagtttg tacaaatgat gacagcgaag ggtggtcccg gggcagcaaa cgacaaaact
     1861 acgctttagc agcttagaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     1921 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     1981 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     2041 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     2101 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2161 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     2221 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     2281 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     2341 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2401 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2461 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2521 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2581 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2641 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2701 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2761 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2821 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2881 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2941 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3001 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3061 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3541 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4141 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4201 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     4261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     4321 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4381 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4441 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
     4501 cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg
     4561 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac
     4621 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
     4681 ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga
     4741 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
     4801 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
     4861 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
     4921 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
     4981 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc
     5041 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
     5101 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
     5161 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
     5221 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
     5281 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     5341 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     5401 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     5461 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagag
//
