LOCUS       sequence-362761-        4857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear-localized LgBiT-CaM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4857)
  AUTHORS   Erdenee E, Ting AY
  TITLE     A Dual-Purpose Real-Time Indicator and Transcriptional Integrator 
            for Calcium Detection in Living Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1086-1095. doi: 
            10.1021/acssynbio.1c00597. Epub 2022 Mar 7.
  PUBMED    35254056
REFERENCE   2  (bases 1 to 4857)
  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: 
            "10.1021/acssynbio.1c00597"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1086-1095"
COMMENT     Sequence Label: NLS-LgBiT-CaM
FEATURES             Location/Qualifiers
     source          1..4857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             827..847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1379..1402
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1878..1895)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1884..2091
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2120..2260
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2120..2249
                     /label=AAV2 ITR
     rep_origin      2335..2790
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2422..2441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2632..2653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2807..2826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2926..2948
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2986..3004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3072..3176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3177..3245,3246..4037)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3395..3414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4208..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4697..4716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgaagcgga ctgctgatgg cagtgaattt gagagcccca aaaagaagag
      841 gaaagtggtc ttcactctgg aagatttcgt aggtgactgg gagcaaacag cagcatacaa
      901 tcttgatcaa gtgttggagc aagggggggt aagctctctc ctccaaaact tggctgtctc
      961 agttactcca attcaacgaa ttgtacggtc aggggaaaat gcactcaaga ttgacattca
     1021 tgtcatcatt ccttatgagg gtctgagtgc cgatcagatg gcccagatcg aggaggtttt
     1081 taaggtagtg tatcccgtgg atgatcatca ctttaaggta attttgccct acggaactct
     1141 ggtaattgac ggagtgaccc ccaatatgct caattatttt ggtaggccct atgaaggtat
     1201 tgctgtcttc gacggaaaaa aaataacagt tactgggaca ctctggaatg gtaataagat
     1261 tatagacgaa cgcttgatca ctcctgacgg cagtatgttg ttccgcgtta ccataaatgg
     1321 gtctgggaac ggaaatggca atggtaacgg gaatggaaac ggcaatggta acgggaatga
     1381 ctacaaggat gacgacgata aggaccaact gactgaagag cagatcgcag aatttaaaga
     1441 ggctttctcc ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac
     1501 ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga
     1561 agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag
     1621 aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg tgtttgataa
     1681 ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga
     1741 gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg
     1801 tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aagtaggaat tcctcgagct
     1861 agagcctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1921 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1981 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2041 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg aggtaaccac
     2101 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2161 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2221 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     2281 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2341 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2401 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2461 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2521 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2581 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2641 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     2701 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2761 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     2821 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2881 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2941 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3001 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3061 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3721 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
     3901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
