LOCUS       sequence-365819-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of recombinant beta1 integrin cytosolic domain in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  AUTHORS   Case LB, De Pasquale M, Henry L, Rosen MK
  TITLE     Synergistic phase separation of two pathways promotes integrin 
            clustering and nascent adhesion formation.
  JOURNAL   Elife. 2022 Jan 20;11. pii: 72588. doi: 10.7554/eLife.72588.
  PUBMED    35049497
REFERENCE   2  (bases 1 to 6894)
  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: "Elife."; 
            date: "2022-01-20"; volume: "11. pii: 72588. doi"; pages: " 
            10.7554/eLife.72588"
COMMENT     Sequence Label: B1 Integrin
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..34
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(28..219)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        715..792
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             793..1875
                     /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(812..831)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2118..2123,2124..2139,2140..2146)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2154..2170
                     /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)."
     primer_bind     2159..2181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2240..3340
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTLEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      3197..3206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3314..3337
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3389..3400
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3413..3433
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3644..3661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3674..3691)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3674..3690)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3683..3705)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3869..3886)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4039..4125
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4217..4244
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4263..4354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4355..4423,4424..5215)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4573..4592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5257..5770)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5452..5473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5664..5683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      5881..6469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6370..6389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6623..6640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6746..6765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
       61 gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga tgtctgcctg
      121 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
      181 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
      241 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
      301 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
      361 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
      421 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
      481 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
      541 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
      601 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
      661 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaaa ttccgacacc
      721 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
      781 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
      841 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
      901 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
      961 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     1021 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     1081 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     1141 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     1201 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     1261 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     1321 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     1381 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     1441 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     1501 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     1561 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     1621 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     1681 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     1741 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     1801 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     1861 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacaatt
     1921 ctcatgtttg acagcttatc atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     1981 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     2041 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     2101 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     2161 cggataacaa tttcacacag gaaacagcca gtccgtttag gtgttttcac gagcacttca
     2221 ccaacaagga ccatagatta tgaaaactga agaaggtaaa ctggtaatct ggattaacgg
     2281 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
     2341 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
     2401 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
     2461 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
     2521 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
     2581 atcgctgatt tataacaaag acctgctgcc gaacccgcca aaaaccttgg aagagatccc
     2641 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
     2701 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
     2761 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
     2821 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
     2881 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
     2941 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     3001 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     3061 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     3121 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa
     3181 agatccacgt attgccgcca ccatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     3241 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     3301 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct cgaacaacaa
     3361 caacaataac aataacaaca acctcgggat cgagggaagg atttcagaat ttgaaaacct
     3421 gtattttcag ggcggttcgc tcgagtgtaa actgcttatg atcattcatg atcgccgcga
     3481 atttgccaaa tttgagaagg aaaaaatgaa cgctaaatgg gatacgggcg aaaaccccat
     3541 ttacaagtca gctgtgacga ccgttgtaaa cccgaagtat gaggggaaac tgcctgaaac
     3601 aggataatag ggatcctcta gagtcgacct gcaggcaagc ttgcatcacc accatcacca
     3661 ttaatagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     3721 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     3781 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cagcttggct
     3841 gttttggcgg atgagataag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     3901 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     3961 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     4021 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     4081 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     4141 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     4201 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     4261 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgtttcccaa tgatgagcac
     4561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5221 agaccaagtt tactcatata tactttagat tgatttaccc cggttgataa tcagaaaagc
     5281 cccaaaaaca ggaagattgt ataagcaaat atttaaattg taaacgttaa tattttgtta
     5341 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5401 aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg
     5461 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5521 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     5581 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5641 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5701 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5761 cagggcgcgt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6001 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6541 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6601 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     6661 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatgtggt gcactctcag
     6721 tacaccgcat gtggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6781 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     6841 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagc
//
