LOCUS       sequence-362767-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear-localized LgBiT-GAI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  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 4692)
  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-GAI
FEATURES             Location/Qualifiers
     source          1..4692
                     /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"
     CDS             1415..1432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1713..1730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1719..1926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1955..2095
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1955..2084
                     /label=AAV2 ITR
     rep_origin      2170..2625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2257..2276)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2467..2488
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2642..2661)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2761..2783
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2821..2839)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2907..3011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3012..3080,3081..3872)
                     /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(3230..3249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4043..4631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4532..4551
                     /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 agatgaagag agatcatcat catcatcatc atcaagataa
     1441 gaagactatg atgatgaatg aagaagacga cggtaacggc atggatgagc ttctagctgt
     1501 tcttggttac aaggttaggt catccgaaat ggctgatgtt gctcagaaac tcgagcagct
     1561 tgaagttatg atgtctaatg ttcaagaaga cgatctttct caactcgcta ctgagactgt
     1621 tcactataat ccggcggagc tttacacgtg gcttgattct atgctcaccg accttaatta
     1681 ggaattcctc gagctagagc ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     1741 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1801 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1861 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1921 tggggaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     1981 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2041 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2101 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     2161 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2221 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     2281 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2341 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     2401 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2461 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     2521 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     2581 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     2641 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2701 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2761 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     2821 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2881 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2941 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3001 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3061 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3121 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3181 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3241 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3301 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3361 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3421 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3481 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3541 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3601 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3661 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3721 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3781 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3841 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3901 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3961 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4021 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4681 ttgctcacat gt
//
