LOCUS       sequence-39157-e        6706 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   Ding Y, Ai HW, Hoi H, Campbell RE
  TITLE     Forster resonance energy transfer-based biosensors for 
            multiparameter ratiometric imaging of Ca2+ dynamics and caspase-3 
            activity in single cells.
  JOURNAL   Anal Chem. 2011 Dec 15;83(24):9687-93. doi: 10.1021/ac202595g. Epub 
            2011 Nov 28.
  PUBMED    22080726
REFERENCE   2  (bases 1 to 6706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/ac202595g";
            journalName: "Anal Chem"; date: "2011-12-15- 15"; volume: "83"; 
            issue: "24"; pages: "9687-93"
COMMENT     Sequence Label: CaYin1
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..284
                     /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             311..333
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             353..370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             374..406
                     /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             419..439
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2387..2408)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3107..3124)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3277..3363
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3455..3482
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3501..3592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3593..3661,3662..4453)
                     /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(3811..3830)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4624..5212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5113..5132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5366..5383
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5398..5538
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5464..5483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5624..5646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5802..6680)
                     /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 tcgcactctc tactgtttct ccatacccgt ttttttgggc
      301 tagaaataat tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca
      361 tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga
      421 gaacctgtac ttccagggct cgagcatggt gagcaagggc gaggaggtca tcaaagagtt
      481 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
      541 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      601 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
      661 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
      721 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
      781 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
      841 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
      901 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
      961 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     1021 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     1081 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     1141 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa
     1201 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     1261 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1321 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     1381 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     1441 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     1501 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     1561 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     1621 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     1681 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     1741 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     1801 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgc gcatgcatct
     1861 gacagaagag cagattgcag agttcaaaga agccttctca ttattggaca aggatgggga
     1921 cggcaccatc accacaaagg aacttggcac cgctctgagg tcgcttggac aaaacccaac
     1981 ggaagcagaa ttgcaggata tgatcaatga agtcgatgct gatggcaatg gaacgattta
     2041 ctttcctgaa tttcttacta tgatggctag aaaaatgaag gacacagaca gcgaagagga
     2101 aatccgagaa gcattccgtg tttttgacaa ggatgggaac ggctacatca gcgctgctga
     2161 attacgtcac gtcatgacaa acctcgggga gaagttaaca gatgaagaag ttgatgaaat
     2221 gataagggaa gcagatatcg atggtgatgg ccaagtaaac tatgaagagt ttgtacaaat
     2281 gatgacagca aaggggggga agaggcgctg gcagaaaaca ggccatgctg tccgagcatt
     2341 cggaagactg aagaagatcg agctcttcac cggggtggtg cccatcctgg tcgagctgga
     2401 cggcgacgta aacggccaca ggttcagcgt gcgcggcgag ggcgagggcg atgccaccaa
     2461 cggcaagctg atcctgaagt tcatctgcac ctccggcgag ctgcccgtgc cctggcccac
     2521 cctcgtgacc accctgtctt acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     2581 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2641 cttcaaggac gacggcagct acaggacccg cgccgaggtg aagttcgagg gcgacaccct
     2701 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2761 caagctggag tacaacatga acgcgtggga cgcgtatatc acggccgaca agcagaagaa
     2821 cggcatcaaa gcgaacttca agatcgagca caacgtcaag gacggcggcg tgcagctcgc
     2881 cgacgcgtac cagcagaaca cccccatcgg cggcggcccc gtgctgctgc ctgacaacca
     2941 ctacctgagc ttccagagca agctgttcaa agacccctac gagcagcgcg atcacatggt
     3001 cctgctggag ttcgttaccg ccgccgggat aactcccggc atggacgagc tgtacaagta
     3061 agaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     3121 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     3181 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     3241 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     3301 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     3361 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     3421 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     3481 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3721 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3781 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3841 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3901 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3961 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4021 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4081 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4141 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4201 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4261 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4321 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4381 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4441 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4501 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4561 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4621 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4681 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4741 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4801 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4861 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4921 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4981 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5041 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5101 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5161 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5221 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5281 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5341 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     5401 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     5461 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     5521 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5581 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5641 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag
     5701 gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta
     5761 tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct
     5821 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat
     5881 tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg
     5941 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg
     6001 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc
     6061 gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg
     6121 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta
     6181 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa
     6241 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc
     6301 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc
     6361 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc
     6421 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt
     6481 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg
     6541 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc
     6601 tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga
     6661 tcattttgcg cttcagccat acttttcata ctcccgccat tcagag
//
