LOCUS       sequence-357183-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human fibronectin fragment, type-III repeats 8-10, R1379A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  AUTHORS   Su Y, Iacob RE, Li J, Engen JR, Springer TA
  TITLE     Dynamics of integrin alpha5beta1, fibronectin, and their complex 
            reveal sites of interaction and conformational change.
  JOURNAL   J Biol Chem. 2022 Sep;298(9):102323. doi: 10.1016/j.jbc.2022.102323.
            Epub 2022 Aug 2.
  PUBMED    35931112
REFERENCE   2  (bases 1 to 6516)
  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: "J Biol 
            Chem."; date: "2022-09"; volume: "298(9):102323. doi"; pages: " 
            10.1016/j.jbc.2022.102323. Epub 2022 Aug 2"
COMMENT     Sequence Label: FnR8-10
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     969..988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1008..1085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1394..1413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1394..1412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1413..1437
                     /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)."
     RBS             1452..1474
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1485..1511
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             1512..1532
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2410..2428)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2424..2471
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2646..2651,2652..2668,2669..2674))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2701..2719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2787..2891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2892..2960,2961..3752)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3110..3129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3923..4511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4412..4431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4665..4682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4697..4836
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4762..4781)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4922..4944
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4938..5129)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6441..6516,1..1007))
                     /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"
ORIGIN
        1 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
       61 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      121 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      181 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
      241 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      301 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      361 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      421 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      481 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      541 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      601 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      661 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      721 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      781 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
      841 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
      901 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
      961 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1021 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1081 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     1141 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     1381 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     1441 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccac caccaccatc
     1501 atcaccatca cgaaaactta tacttccagg gtgttcctcc tcccactgac ctgcgattca
     1561 ccaacattgg tccagacacc atgcgtgtca cctgggctcc acccccatcc attgatttaa
     1621 ccaacttcct ggtgcgttac tcacctgtga aaaatgagga agatgttgca gagttgtcaa
     1681 tttctccttc agacaatgca gtggtcttaa caaatctcct gcctggtaca gaatatgtag
     1741 tgagtgtctc cagtgtctac gaacaacatg agagcacacc tcttagagga agacagaaaa
     1801 caggtcttga ttccccaact ggcattgact tttctgatat tactgccaac tcttttactg
     1861 tgcactggat tgctcctcga gccaccatca ctggctacag gatccgccat catcccgagc
     1921 acttcagtgg gagacctcga gaagatcggg tgccccactc tgcgaattcc atcaccctca
     1981 ccaacctcac tccaggcaca gagtatgtgg tcagcatcgt tgctcttaat ggcagagagg
     2041 aaagtccctt attgattggc caacaatcaa cagtttctga tgttccgagg gacctggaag
     2101 ttgttgctgc gacccccacc agcctactga tcagctggga tgctcctgct gtcacagtga
     2161 gatattacag gatcacttac ggagagacag gaggaaatag ccctgtccag gagttcactg
     2221 tgcctgggag caagtctaca gctaccatca gcggccttaa acctggagtt gattatacca
     2281 tcactgtgta tgctgtcact ggccgtggag acagccccgc aagcagcaag ccaatttcca
     2341 ttaattaccg aacatgatag gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     2401 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     2461 ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc ccggcagtac
     2521 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
     2581 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
     2641 accgcattaa agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg
     2701 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2761 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2821 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2881 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2941 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3001 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3061 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3121 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3181 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3241 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3301 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3361 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3421 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3481 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3541 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3601 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3661 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3721 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3781 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3841 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3901 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3961 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4021 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4081 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4141 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4201 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4261 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4321 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4381 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4441 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4501 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4561 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4621 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4681 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4741 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4801 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4861 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4921 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4981 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     5041 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     5101 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     5161 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     5221 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     5281 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     5341 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     5401 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     5461 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     5521 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     5581 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     5641 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     5701 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     5761 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     5821 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     5881 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
     5941 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     6001 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     6061 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     6121 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     6181 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     6241 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6301 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6361 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     6421 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6481 tgcattaatg aatcggccaa cgcgcgggga gaggcg
//
