LOCUS       sequence-347674-        2737 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mir218 binding site2 point mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2737)
  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 2737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_1745G>U
FEATURES             Location/Qualifiers
     source          1..2737
                     /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(561..577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(561..577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(574..596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    585..601
                     /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(609..615,616..633,634..639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    654..675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(792..809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(963..1551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1043..1062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1722..2537)
                     /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     2445..2464
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2538..2642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2710..2728
                     /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 gggatggaac
      421 cagtggaact tcagctggac tggcagaagc agaagggagg ggtacggcgg tcactgttcc
      481 tggcctccag gcaggcaacc ctgacccaga ccctgctcat cgttccatcc caatggcgcg
      541 ccgagcttgg ctcgagcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
      601 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
      661 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
      721 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      781 tgttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttagaaaaa ctcatcgagc
     1741 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     1801 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     1861 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     1921 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     1981 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2041 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gaaacgaaat
     2101 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2161 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2221 gctgttttcc cagggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     2281 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     2341 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     2401 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     2461 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     2521 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     2581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2641 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2701 acctataaaa ataggcgtat cacgaggccc ttttgtc
//
