LOCUS       sequence-319308-        7668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains integrin alpha v residues 1–594 with the M400C mutation and
            Gly inserted prior to residue 400.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7668)
  AUTHORS   Dong X, Hudson NE, Lu C, Springer TA
  TITLE     Structural determinants of integrin beta-subunit specificity for 
            latent TGF-beta.
  JOURNAL   Nat Struct Mol Biol. 2014 Dec;21(12):1091-6. doi: 10.1038/nsmb.2905.
            Epub 2014 Nov 10.
  PUBMED    25383667
REFERENCE   2  (bases 1 to 7668)
  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: "10.1038/nsmb.2905";
            journalName: "Nat Struct Mol Biol"; date: "2014-12"; volume: "21"; 
            issue: "12"; pages: "1091-6"
COMMENT     Sequence Label: Integrin-alpha-v Headpiece M400GC
FEATURES             Location/Qualifiers
     source          1..7668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     771..790
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        771..789
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2720..2743
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2807..2818
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2951..2968
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2998..3015)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3004..3228
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3274..3702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3361..3380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3571..3592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3711..3731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3716..4045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3896..4031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3958..3977
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4094..5119
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4094..5119
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5249..5370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5286..5305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5340..5359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5419..5435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5419..5435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5432..5454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5443..5459
                     /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)."
     promoter        complement(join(5467..5473,5474..5491,5492..5497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5512..5533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5650..5667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5821..6409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5901..5920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6580..7371,7372..7440))
                     /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     7203..7222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7441..7545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7620..7639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      781 cactataggg agacccaagc tggctagcgt ttaaacgggc cctctagagg cgatggcttt
      841 tccgccgcgg cgacggctgc gcctcggtcc ccgcggcctc ccgcttcttc tctcgggact
      901 cctgctacct ctgtgccgcg ccttcaacct agacgtggac agtcctgccg agtactctgg
      961 ccccgaggga agttacttcg gcttcgccgt ggatttcttc gtgcccagcg cgtcttcccg
     1021 gatgtttctt ctcgtgggag ctcccaaagc aaacaccacc cagcctggga ttgtggaagg
     1081 agggcaggtc ctcaaatgtg actggtcttc tacccgccgg tgccagccaa ttgaatttga
     1141 tgcaacaggc aatagagatt atgccaagga tgatccattg gaatttaagt cccatcagtg
     1201 gtttggagca tctgtgaggt cgaaacagga taaaattttg gcctgtgccc cattgtacca
     1261 ttggagaact gagatgaaac aggagcgaga gcctgttgga acatgctttc ttcaagatgg
     1321 aacaaagact gttgagtatg ctccatgtag atcacaagat attgatgctg atggacaggg
     1381 attttgtcaa ggaggattca gcattgattt tactaaagct gacagagtac ttcttggtgg
     1441 tcctggtagc ttttattggc aaggtcagct tatttcggat caagtggcag aaatcgtatc
     1501 taaatacgac cccaatgttt acagcatcaa gtataataac caattagcaa ctcggactgc
     1561 acaagctatt tttgatgaca gctatttggg ttattctgtg gctgtcggag atttcaatgg
     1621 tgatggcata gatgactttg tttcaggagt tccaagagca gcaaggactt tgggaatggt
     1681 ttatatttat gatgggaaga acatgtcctc cttatacaat tttactggcg agcagatggc
     1741 tgcatatttc ggattttctg tagctgccac tgacattaat ggagatgatt atgcagatgt
     1801 gtttattgga gcacctctct tcatggatcg tggctctgat ggcaaactcc aagaggtggg
     1861 gcaggtctca gtgtctctac agagagcttc aggagacttc cagacgacaa agctgaatgg
     1921 atttgaggtc tttgcacggt ttggcagtgc catagctcct ttgggagatc tggaccagga
     1981 tggtttcaat gatattgcaa ttgctgctcc atatgggggt gaagataaaa aaggaattgt
     2041 ttatatcttc aatggaagat caacaggctt gaacgcagtc ccatctcaaa tccttgaagg
     2101 gcagtgggct gctcgaagcg gatgcccacc aagctttggc tattcaatga aaggagccac
     2161 agatatagac aaaaatggat atccagactt aattgtagga gcttttggtg tagatcgagc
     2221 tatcttatac agggccagac cagttatcac tgtaaatgct ggtcttgaag tgtaccctag
     2281 cattttaaat caagacaata aaacctgctc actgcctgga acagctctca aagtttcctg
     2341 ttttaatgtt aggttctgct taaaggcaga tggcaaagga gtacttccca ggaaacttaa
     2401 tttccaggtg gaacttcttt tggataaact caagcaaaag ggagcaattc gacgagcact
     2461 gtttctctac agcaggtccc caagtcactc caagaacatg actatttcaa gggggggact
     2521 gatgcagtgt gaggaattga tagcgtatct gcgggatgaa tctgaattta gagacaaact
     2581 cactccaatt actattttta tggaatatcg gttggattat agaacagctg ctgatacaac
     2641 aggcttgcaa cccattctta accagttcac gcctgctaac attagtcgac aggctcacat
     2701 tctacttgac accggtggac tggaggtgct cttccaaggc cccggagaaa atgctcagct
     2761 ggaaaaagag ctccaggcac ttgagaagga aaatgcacag ctggaatggg aattacaagc
     2821 actagagaag gagctagctc aaaccactgg ttggagaggt ggccatgttg tggaaggctt
     2881 ggctggtgaa ttggaacaat tgagagcccg tctggagcat cacccacaag gccaaagaga
     2941 accagcgggc catcaccatc accatcacta aaagcttaag tttaaaccgc tgatcagcct
     3001 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3061 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3121 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3181 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3241 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3481 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3601 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3721 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3781 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3841 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3901 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3961 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4021 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4081 tgatcagcac gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4141 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4201 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4261 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4321 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4381 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4441 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4501 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4561 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4621 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4681 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4741 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4801 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4861 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4921 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4981 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     5041 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5101 tccgagggca aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga
     5161 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5221 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5341 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5401 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5461 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7621 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgct
//
