LOCUS       sequence-347672-        2731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mir218 binding site1 point mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2731)
  AUTHORS   Liu J, Li Y, Lyu L, Xiao L, Memon AA, Yu X, Halim A, Patel S, Osman 
            A, Yin W, Jiang J, Naini S, Lim K, Zhang A, Williams JD, Koester R, 
            Qi KZ, Fucci QA, Ding L, Chang S, Patel A, Mori Y, Chaudhari A, Bao 
            A, Liu J, Lu TS, Siedlecki A
  TITLE     Integrin alpha5 Is Regulated by miR-218-5p in Endothelial Progenitor
            Cells.
  JOURNAL   J Am Soc Nephrol. 2022 Mar;33(3):565-582. doi: 
            10.1681/ASN.2021020140. Epub 2022 Jan 28.
  PUBMED    35091451
REFERENCE   2  (bases 1 to 2731)
  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: 
            "10.1681/ASN.2021020140"; journalName: "J Am Soc Nephrol"; date: 
            "2022-03"; volume: "33"; issue: "3"; pages: "565-582"
COMMENT     Sequence Label: pITGA5_473G>A
FEATURES             Location/Qualifiers
     source          1..2731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(555..571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(555..571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(568..590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    579..595
                     /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(603..609,610..627,628..633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    648..669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(786..803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(957..1545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1037..1056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1716..2531)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRFAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2439..2458
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2532..2636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2704..2722
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagactgtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggatggtcc
      421 tggagtcctc actgtccagc tcagagggag aggagcctgt ggagtactag agggagagga
      481 gcctgtggag tacaagtcct tgcagtggtt cggggcaagg atcccaatgg cgcgccgagc
      541 ttggctcgag catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      601 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      661 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      721 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctgttcc
      781 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      841 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      901 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      961 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1021 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1081 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1141 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1201 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1261 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1321 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1381 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1441 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1501 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1561 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1621 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1681 atctaaagta tatatgagta aacttggtct gacagttaga aaaactcatc gagcatcaaa
     1741 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1801 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1861 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1921 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1981 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2041 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgaaacg aaatacgcga
     2101 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2161 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2221 ttcccaggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2281 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2341 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2401 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2461 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2521 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2641 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     2701 aaaaataggc gtatcacgag gcccttttgt c
//
