LOCUS       sequence-496373-        7306 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Mammalian expression vector of human ITGB5 fused with Halo9-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7306)
  AUTHORS   Nishimura R, Barnett SFH, Jain K, Huang Z, Goult BT, Kanchanawong P
  TITLE     Optogenetic control of mechanotransduction based on light-induced 
            homodimerization of talin.
  JOURNAL   J Cell Sci. 2025 Nov 24:jcs.264110. doi: 10.1242/jcs.264110.
  PUBMED    41277405
REFERENCE   2  (bases 1 to 7306)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell Sci.
            2025 Nov 24:jcs.264110. doi: 10.1242/jcs.264110."
COMMENT     Sequence Label: pEGFP-N1/Integrin-b5-FL-SGx3-Halo9
FEATURES             Location/Qualifiers
     source          1..7306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3078..3968
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DHNVFIEGTLRMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    4094..4215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4131..4150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4185..4204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4222..4677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4359..4380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4571..4590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4704..4808
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4810..5167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4810..5001
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4833..4853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5018..5153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5202..5996
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5202..5996
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5256..5275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5866..5885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6184..6203)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6228..6275
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6604..7192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7093..7112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgc
      601 cgcgggcccc ggcgccgctg tacgcctgcc tcctggggct ctgcgcgctc ctgccccggc
      661 tcgcaggtct caacatatgc actagtggaa gtgccacctc atgtgaagaa tgtctgctaa
      721 tccacccaaa atgtgcctgg tgctccaaag aggacttcgg aagcccacgg tccatcacct
      781 ctcggtgtga tctgagggca aaccttgtca aaaatggctg tggaggtgag atagagagcc
      841 cagccagcag cttccatgtc ctgaggagcc tgcccctcag cagcaagggt tcgggctctg
      901 caggctggga cgtcattcag atgacaccac aggagattgc cgtgaacctc cggcccggtg
      961 acaagaccac cttccagcta caggttcgcc aggtggagga ctatcctgtg gacctgtact
     1021 acctgatgga cctctccctg tccatgaagg atgacttgga caatatccgg agcctgggca
     1081 ccaaactcgc ggaggagatg aggaagctca ccagcaactt ccggttggga tttgggtctt
     1141 ttgttgataa ggacatctct cctttctcct acacggcacc gaggtaccag accaatccgt
     1201 gcattggtta caagttgttt ccaaattgcg tcccctcctt tgggttccgc catctgctgc
     1261 ctctcacaga cagagtggac agcttcaatg aggaagttcg gaaacagagg gtgtcccgga
     1321 accgagatgc ccctgagggg ggctttgatg cagtactcca ggcagccgtc tgcaaggaga
     1381 agattggctg gcgaaaggat gcactgcatt tgctggtgtt cacaacagat gatgtgcccc
     1441 acatcgcatt ggatggaaaa ttgggaggcc tggtgcagcc acacgatggc cagtgccacc
     1501 tgaacgaggc caacgagtac actgcatcca accagatgga ctatccatcc cttgccttgc
     1561 ttggagagaa attggcagag aacaacatca acctcatctt tgcagtgaca aaaaaccatt
     1621 atatgctgta caagaatttt acagccctga tacctggaac aacggtggag attttagatg
     1681 gagactccaa aaatattatt caactgatta ttaatgcata caatagtatc cggtctaaag
     1741 tggagttgtc agtctgggat cagcctgagg atcttaatct cttctttact gctacctgcc
     1801 aagatggggt atcctatcct ggtcagagga agtgtgaggg tctgaagatt ggggacacgg
     1861 catcttttga agtatcattg gaggcccgaa gctgtcccag cagacacacg gagcatgtgt
     1921 ttgccctgcg gccggtggga ttccgggaca gcctggaggt gggggtcacc tacaactgca
     1981 cgtgcggctg cagcgtgggg ctggaaccca acagcgccag gtgcaacggg agcgggacct
     2041 atgtctgcgg cctgtgtgag tgcagccccg gctacctggg caccaggtgc gagtgccagg
     2101 atggggagaa ccagagcgtg taccagaacc tgtgccggga ggcagagggc aagccactgt
     2161 gcagcgggcg tggggactgc agctgcaacc agtgctcctg cttcgagagc gagtttggca
     2221 agatctatgg gcctttctgt gagtgcgaca acttctcctg tgccaggaac aagggagtcc
     2281 tctgctcagg ccatggcgag tgtcactgcg gggaatgcaa gtgccatgca ggttacatcg
     2341 gggacaactg taactgctcg acagacatca gcacatgccg gggcagagat ggccagatct
     2401 gcagcgagcg tgggcactgt ctctgtgggc agtgccaatg cacggagccg ggggcctttg
     2461 gggagatgtg tgagaagtgc cccacctgcc cggatgcatg cagcaccaag agagattgcg
     2521 tcgagtgcct gctgctccac tctgggaaac ctgacaacca gacctgccac agcctatgca
     2581 gggatgaggt gatcacatgg gtggacacca tcgtgaaaga tgaccaggag gctgtgctat
     2641 gtttctacaa aaccgccaag gactgcgtca tgatgttcac ctatgtggag ctccccagtg
     2701 ggaagtccaa cctgaccgtc ctcagggagc cagagtgtgg aaacaccccc aacgccatga
     2761 ccatcctcct ggctgtggtc ggtagcatcc tccttgttgg gcttgcactc ctggctatct
     2821 ggaagctgct tgtcaccatc cacgaccgga gggagtttgc aaagtttcag agcgagcgat
     2881 ccagggcccg ctatgaaatg gcttcaaatc cattatacag aaagcctatc tccacgcaca
     2941 ctgtggactt caccttcaac aagttcaaca aatcctacaa tggcactgtg gacaagcttc
     3001 gaattctgca gtcgacggta ccgcgggatc catctggtgg cggagggagt ggcggaggtg
     3061 ggtcaggagg tggaggcatg gcagaaatcg gtactggctt tccattcgac ccccattatg
     3121 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
     3181 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
     3241 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
     3301 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
     3361 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
     3421 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     3481 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     3541 ccgacgtcgg ccgcaagctg atcatcgatc acaacgtttt tatcgagggt acgctgcgga
     3601 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     3661 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     3721 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     3781 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     3841 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     3901 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg acgctcgaga
     3961 tttccggcta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     4021 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4081 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4141 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4201 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4261 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4321 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4381 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4441 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4501 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4561 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4621 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4801 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5101 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5161 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5221 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5281 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5341 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5401 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5461 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5521 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5581 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5641 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5701 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5761 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5821 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5881 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5941 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6001 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6061 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6181 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6241 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6301 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6361 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6421 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6481 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6541 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6601 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6661 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6721 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6781 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6841 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6901 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6961 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7021 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7081 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7141 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7201 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7261 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
