LOCUS       sequence-384233-        6698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent labeling for cell membranes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  AUTHORS   Hsu YY, Resto Irizarry AM, Fu J, Liu AP
  TITLE     Mechanosensitive Channel-Based Optical Membrane Tension Reporter.
  JOURNAL   ACS Sens. 2023 Jan 27;8(1):12-18. doi: 10.1021/acssensors.2c01921. 
            Epub 2023 Jan 6.
  PUBMED    36608338
REFERENCE   2  (bases 1 to 6698)
  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.2c01921"; journalName: "ACS Sens"; date: 
            "2023-01-27- 27"; volume: "8"; issue: "1"; pages: "12-18"
COMMENT     Sequence Label: Halo-TMD
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      773..859
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      951..978
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     986..1009
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    1028..1127
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        1650..2029
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2030..2233
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2184..2204
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2235..2253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2256..2274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2284..2308
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          2423..3249
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     promoter        complement(4501..4519)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4502..4519)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4539..4556)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4545..4769
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4831..4930)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5042..5061)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5053..5862
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5120..5139)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(join(6357..6698,1..19))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6371..6509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(6526..6698,1..19)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6674..6694
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ctggttcttt ccgcctcagt tgagatcctt tttttctgcg tgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca
      721 tggatctcgg ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa
      781 acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      841 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg
      901 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat
      961 cctgacggat ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct
     1021 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     1081 taagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac
     1141 ccgcgttttg agatttctgt cgccgactaa attcatgtcg cgcgatagtg gtgtttatcg
     1201 ccgatagaga tggcgatatt ggaaaaatcg atatttgaaa atatggcata ttgaaaatgt
     1261 cgccgatgtg agtttctgtg taactgatat cgccattttt ccaaaagtga tttttgggca
     1321 tacgcgatat ctggcgatag cgcttatatc gtttacgggg gatggcgata gacgactttg
     1381 gtgacttggg cgattctgtg tgtcgcaaat atcgcagttt cgatataggt gacagacgat
     1441 atgaggctat atcgccgata gaggcgacat caagctggca catggccaat gcatatcgat
     1501 ctatacattg aatcaatatt ggccattagc catattattc attggttata tagcataaat
     1561 caatattggc tattggccat tgcatacgtt gtatccatat cataatatgt acatttatat
     1621 tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta
     1681 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1741 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1801 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1861 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1921 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1981 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2041 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2101 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2161 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2221 tataagcaga gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg
     2281 acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     2341 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
     2401 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
     2461 ggcttcttat gcatgctata ctgtttttgg cttggggtct atacaccccc gcttcctcat
     2521 gttataggtg atggtatagc ttagcctata ggtgtgggtt attgaccatt attgaccact
     2581 cccctattgg tgacgatact ttccattact aatccataac atggctcttt gccacaactc
     2641 tctttattgg ctatatgcca atacactgtc cttcagagac tgacacggac tctgtatttt
     2701 tacaggatgg ggtctcattt attatttaca aattcacata tacaacacca ccgtccccag
     2761 tgcccgcagt ttttattaaa cataacgtgg gatctccacg cgaatctcgg gtacgtgttc
     2821 cggacatggg ctcttctccg gtagcggcgg agcttctaca tccgagccct gctcccatgc
     2881 ctccagcgac tcatggtcgc tcggcagctc cttgctccta acagtggagg ccagacttag
     2941 gcacagcacg atgcccacca ccaccagtgt gccgcacaag gccgtggcgg tagggtatgt
     3001 gtctgaaaat gagctcgggg agcgggcttg caccgctgac gcatttggaa gacttaaggc
     3061 agcggcagaa gaagatgcag gcagctgagt tgttgtgttc tgataagagt cagaggtaac
     3121 tcccgttgcg gtgctgttaa cggtggaggg cagtgtagtc tgagcagtac tcgttgctgc
     3181 cgcgcgcgcc accagacata atagctgaca gactaacaga ctgttccttt ccatgggtct
     3241 tttctgcagt caccgtcctt gacacgaagc ttggtaccgc caccatgatg gatcaagcta
     3301 gatcagcatt ctctaacttg tttggtggag aaccattgtc atgtacccgg gccagcctgg
     3361 ctcggcaagt agatggcgat aacagtcatg tggagatgaa acttgctgta gatgaagaag
     3421 aaaatgctga caataacaca aaggccaatg tcacaaaacc aaaaaggtgt agtggaagta
     3481 tctgctatgg gactattgct gtgatcgtct ttttcttgat tggatttatg attggctact
     3541 tgggctatgg aagcggagga tccgggatcg gcacgggctt tccgtttgac ccacactatg
     3601 tcgaagtgct tggcgaacgt atgcattacg tggacgtcgg tcctcgggac ggtaccccag
     3661 tcctcttctt acacggcaat cctacatctt cttatgtgtg gcgtaatatc attccacacg
     3721 tcgcaccaac gcaccgctgt atcgcaccgg atctgattgg catggggaag tctgacaagc
     3781 cggacttagg ttatttcttc gacgaccacg tacggttcat ggacgcgttt attgaggcat
     3841 taggtctgga ggaagttgtt ctggtgatcc atgattgggg ttctgcactt ggctttcact
     3901 gggctaagcg gaacccagaa cgtgtgaagg ggatcgcttt tatggagttc attcgcccta
     3961 ttcctacttg ggatgaatgg ccagaatttg cgcgcgagac atttcaagca tttcggacaa
     4021 cggacgtagg tcggaagctc atcattgatc aaaacgtatt catcgagggc acactcccga
     4081 tgggggttgt gcgtccactc acagaagtcg agatggatca ttatcgtgag ccgtttttga
     4141 accctgtgga tcgcgaaccg ctgtggcgtt ttcctaacga gctgccaatt gctggggaac
     4201 cggctaacat cgtggccctt gtcgaagaat acatggactg gttacaccaa agtcctgttc
     4261 caaagttgct cttttggggg acgccggggg tgctgattcc tccggctgaa gcagctcggt
     4321 tggcaaagag tctcccaaat tgcaaagcgg tagatattgg tcctggttta aacttattgc
     4381 aggaggataa tcctgacctt attggctctg agattgcccg gtggctttct acactggaga
     4441 tctccgggct ggaagaattc ctcgaggcgg ccgctcgagc atgcatctag agggccctat
     4501 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4561 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4621 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4681 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4741 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4801 gaggggggat cgatccgtcg agatatctag acccagcttt cttgtacaaa gttggcatta
     4861 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     4921 tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     4981 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     5041 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     5101 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     5161 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     5221 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     5281 atgcctcttc cgaccatcaa gcattttatc cgtactcctg gtgatgcatg gttactcacc
     5341 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     5401 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     5461 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     5521 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     5581 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     5641 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     5701 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     5761 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     5821 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     5881 ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc
     5941 aaaggatctt ctcagtcctg ctcctctgcc acaaagtgca cacagttgcc tgctgggtct
     6001 ctcagggcaa actctctccc ccagggctgc tctccaatct ctgtcatggc agggccactg
     6061 gcatccctga agttggtgga gaccacctca ctccactcag catacagctc atccagtcct
     6121 ctcacccaca cccaagccag ggtgttgtca ggcaccacct ggtcctggac tgctgagatg
     6181 aacagggtga catcatctct gaccacacct gcaaagtcat cctccacaaa atctctggag
     6241 aaccccaacc tgtcagtcca gaactcaaca gctccagcca catccctggc tgtgagcact
     6301 gggacagcac tggtcaactt ggccataacg atcctcatcc tgtctcttga tcgatctttg
     6361 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     6421 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     6481 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     6541 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     6601 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6661 ggggactttc cacaccctaa ctgacacaca ttccacag
//
