LOCUS       sequence-428946-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HaloTag-STReTCh-RGD fusion protein in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5300)
  AUTHORS   Zhong BL, Vachharajani VT, Dunn AR
  TITLE     Facile detection of mechanical forces across proteins in cells with 
            STReTCh.
  JOURNAL   Cell Rep Methods. 2022 Sep 2;2(9):100278. doi: 
            10.1016/j.crmeth.2022.100278. eCollection 2022 Sep 19.
  PUBMED    36160040
REFERENCE   2  (bases 1 to 5300)
  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"; journalName: "Cell Rep 
            Methods."; date: "2022-09-2"; volume: "2(9):100278. doi"; pages: " 
            10.1016/j.crmeth.2022.100278. eCollection 2022 Sep 19"
COMMENT     Sequence Label: T7-LacO-Halo-STReTCh-RGD
FEATURES             Location/Qualifiers
     source          1..5300
                     /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             3144..3182
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             3294..3311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3319..3366
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3589..3608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3614..3633)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3675..3766
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3767..3835,3836..4627)
                     /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(3985..4004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             join(4710..5300,1..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(4729..4748)
                     /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 atgggcagcg aaattggcac tggttttccg tttgatccgc
     2041 attacgttga agttttaggc gaacgcatgc actacgtaga cgtgggtccg cgtgacggca
     2101 ccccggtcct gttcctgcac ggcaatccaa cctctagcta tgtctggcgc aacattatcc
     2161 cgcacgtcgc gccgacgcac cgtagtattg caccggatct gatcggtatg ggtaaaagcg
     2221 ataaaccgga cctgggctat ttctttgacg accacgttcg ttttatggat gcgttcattg
     2281 aggccttggg tctggaagag gtggtgttgg ttatccatga ctggggtagc gctctgggct
     2341 tccactgggc gaagcgcaac ccggaacgtg tgaagggcat cgcatttatg gagttcattc
     2401 gcccgattcc gacctgggac gagtggccgg agttcgcgcg tgaaacgttc caggcctttc
     2461 gcaccaccga tgtcggtcgt aagctgatta ttgaccaaaa tgttttcatc gagggcacgt
     2521 tgccgatggg cgttgttcgt ccgctgaccg aagtcgagat ggaccattac cgtgagccgt
     2581 ttttgaatcc ggtggatcgt gagccgctgt ggcgtttccc gaacgagctg ccgatcgccg
     2641 gtgaaccggc gaacatcgtt gcgctggttg aagagtacat ggactggctg caccagagcc
     2701 cggtgccgaa actgctgttt tggggtacgc cgggtgtgct gatcccgcct gcggaagcag
     2761 cccgcctggc gaaatccctg ccgaacgcga aggcagtgga tattggtcca ggcctgaatc
     2821 tgctgcagga agataacccg gacctgattg gtagcgaaat cgcgcgctgg ctgagcacgt
     2881 tggagatcag cggcggtgct ggcgagttca tggaaactct gcatattacc aaaaccatga
     2941 aaaacattga agtgccggaa accaaaaccg cgagctttga atgcgaagtg agccatttta
     3001 acgtgccgag catgtggctg aaaaacggcg tggaaattga aatgagcgaa aaatttaaaa
     3061 ttgtggtgca gggcaaactg catcagctga ttattatgaa caccagcacc gaagatagcg
     3121 cggaatatac ctttgtgtgc ggcgcgcata ttgtgatggt ggatgcgtat aaaccgacca
     3181 aaaacgatca ggtgagcgcg accctgaccg tgacttccga gaatctgtac ttcgagcaag
     3241 gtaccgtcta tgccgtcacc ggtcgtggcg attccccggc aagctcggcg gcacaccacc
     3301 atcatcacca ttaaccccct agcataaccc cttggggcct ctaaacgggt cttgaggggt
     3361 tttttgcccc tgagacgcgt caatcgagtt cgtacctaag ggcgacaccc cataattagc
     3421 ccgggcgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
     3481 cctactctcg catggggagt ccccacacta ccatcggcgc tacggcgttt cacttctgag
     3541 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca aggggtgtta
     3601 tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt aattggttgt
     3661 aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3721 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga gtattcaaca
     3781 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3841 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3901 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3961 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4021 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4081 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4141 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4201 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4261 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcgatggc
     4321 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4381 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4441 tggctggttt attgctgata aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc
     4501 agcgctgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4561 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4621 ttggtaagcg gcgcgccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     4681 cggaagagag tcaattcagg gtggtgaata tgaaaccagt aacgttatac gatgtcgcag
     4741 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     4801 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     4861 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     4921 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     4981 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     5041 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     5101 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     5161 ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg cgactgggcg
     5221 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     5281 ctgtctcggc gcgtctgcgt
//
