LOCUS       sequence-428952-        4487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LgBiT in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4487)
  AUTHORS   Zhong BL, Elliot JM, Wang P, Li H, Hall RN, Wang B, Prakash M, Dunn 
            AR
  TITLE     Split Luciferase Molecular Tension Sensors for Bioluminescent 
            Readout of Mechanical Forces in Biological Systems.
  JOURNAL   ACS Sens. 2024 Jul 26;9(7):3489-3495. doi: 
            10.1021/acssensors.3c02664. Epub 2024 Jul 7.
  PUBMED    38973210
REFERENCE   2  (bases 1 to 4487)
  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/acssensors.3c02664"; journalName: "ACS Sens"; date: 
            "2024-07-26- 26"; volume: "9"; issue: "7"; pages: "3489-3495"
COMMENT     Sequence Label: T7-LacO-LgBiT
FEATURES             Location/Qualifiers
     source          1..4487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      578..1166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1067..1086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1320..1337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1435..1452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1436..1452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1915..1934
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1915..1933
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1934..1958
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2481..2498
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2506..2553
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2776..2795)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2801..2820)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2862..2953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2954..3022,3023..3814)
                     /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(3172..3191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             join(3897..4487,1..492)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3916..3935)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
       61 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
      121 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
      181 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
      241 gatagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
      301 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
      361 ctgttgccag tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact
      541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      721 ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta
      781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1321 gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa
     1381 gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa
     1441 acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat
     1501 ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag
     1561 agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga gaattattta
     1621 accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga
     1681 tgaacaccta taattaaact attcatctat tatttatgat tttttgtata tacaatattt
     1741 ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt
     1801 tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg
     1861 ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttccgcg aaattaatac
     1921 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     1981 acttttagga ggtaaaacat atggtcttca ctctggaaga tttcgtaggt gactgggagc
     2041 aaacagcagc atacaatctt gatcaagtgt tggagcaagg gggggtaagc tctctcctcc
     2101 aaaacttggc tgtctcagtt actccaattc aacgaattgt acggtcaggg gaaaatgcac
     2161 tcaagattga cattcatgtc atcattcctt atgagggtct gagtgccgat cagatggccc
     2221 agatcgagga ggtttttaag gtagtgtatc ccgtggatga tcatcacttt aaggtaattt
     2281 tgccctacgg aactctggta attgacggag tgacccccaa tatgctcaat tattttggta
     2341 ggccctatga aggtattgct gtcttcgacg gaaaaaaaat aacagttact gggacactct
     2401 ggaatggtaa taagattata gacgaacgct tgatcactcc tgacggcagt atgttgttcc
     2461 gcgttaccat aaatgggtct caccaccatc atcaccatta accccctagc ataacccctt
     2521 ggggcctcta aacgggtctt gaggggtttt ttgcccctga gacgcgtcaa tcgagttcgt
     2581 acctaagggc gacaccccat aattagcccg ggcgaaaggc ccagtctttc gactgagcct
     2641 ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagtccc cacactacca
     2701 tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac caccgcgcta
     2761 ctgccgccag gcaaacaagg ggtgttatga gccatattca ggtataaatg ggctcgcgat
     2821 aatgttcaga attggttaat tggttgtaac actgacccct atttgtttat ttttctaaat
     2881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2941 aaaaaggaag aatatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3061 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3121 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3241 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3361 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3481 tgacaccacg atgcctgtag cgatggcaac aacgttgcgc aaactattaa ctggcgaact
     3541 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ccggagccgg
     3661 tgagcgtggt tctcgcggta tcatcgcagc gctggggcca gatggtaagc cctcccgtat
     3721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3781 tgagataggt gcctcactga ttaagcattg gtaagcggcg cgccatcgaa tggcgcaaaa
     3841 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatatga
     3901 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     3961 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     4021 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     4081 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     4141 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     4201 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     4261 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     4321 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     4381 cccatgagga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     4441 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgt
//
