LOCUS       sequence-232461-        9783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enables eukaryotic expression of human plasma fibronectin in which 
            all three O-glycosylation sites were mutated from threonine to 
            serine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9783)
  AUTHORS   Sens C, Altrock E, Rau K, Klemis V, von Au A, Pettera S, Uebel S, 
            Damm T, Tiwari S, Moser M, Nakchbandi IA
  TITLE     An O-Glycosylation of Fibronectin Mediates Hepatic Osteodystrophy 
            Through alpha4beta1 Integrin.
  JOURNAL   J Bone Miner Res. 2017 Jan;32(1):70-81. doi: 10.1002/jbmr.2916. Epub
            2016 Aug 3.
  PUBMED    27427791
REFERENCE   2  (bases 1 to 9783)
  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: "10.1002/jbmr.2916";
            journalName: "J Bone Miner Res"; date: "2017-01"; volume: "32"; 
            issue: "1"; pages: "70-81"
COMMENT     Sequence Label: Fibronectin-human-mutated N-terminal (N), V1 a...
FEATURES             Location/Qualifiers
     source          1..9783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(7956..7975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    7985..8066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     8010..8029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8264..8852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8344..8363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8933..9727)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(8933..9727)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9044..9063)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9654..9673
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacaggggta ccgccatcat
      961 gaagtttaaa caagcttgaa ttctctagag atatcctgca gagatctgga tccctcgagg
     1021 ctagctgtca acatgcttag gggtccgggg cccgggctgc tgctgctggc cgtccagtgc
     1081 ctggggacag cggtgccctc cacgggagcc tcgaagagca agaggcaggc tcagcaaatg
     1141 gttcagcccc agtccccggt ggctgtcagt caaagcaagc ccggttgtta tgacaatgga
     1201 aaacactatc agataaatca acagtgggag cggacctacc taggcaatgc gttggtttgt
     1261 acttgttatg gaggaagccg aggttttaac tgcgagagta aacctgaagc tgaagagact
     1321 tgctttgaca agtacactgg gaacacttac cgagtgggtg acacttatga gcgtcctaaa
     1381 gactccatga tctgggactg tacctgcatc ggggctgggc gagggagaat aagctgtacc
     1441 atcgcaaacc gctgccatga agggggtcag tcctacaaga ttggtgacac ctggaggaga
     1501 ccacatgaga ctggtggtta catgttagag tgtgtgtgtc ttggtaatgg aaaaggagaa
     1561 tggacctgca agcccatagc tgagaagtgt tttgatcatg ctgctgggac ttcctatgtg
     1621 gtcggagaaa cgtgggagaa gccctaccaa ggctggatga tggtagattg tacttgcctg
     1681 ggagaaggca gcggacgcat cacttgcact tctagaaata gatgcaacga tcaggacaca
     1741 aggacatcct atagaattgg agacacctgg agcaagaagg ataatcgagg aaacctgctc
     1801 cagtgcatct gcacaggcaa cggccgagga gagtggaagt gtgagaggca cacctctgtg
     1861 cagtcctcat cgagcggatc tggccccttc accgatgttc gtgcagctgt ttaccaaccg
     1921 cagcctcacc cccagcctcc tccctatggc cactgtgtca cagacagtgg tgtggtctac
     1981 tctgtgggga tgcagtggct gaagacacaa ggaaataagc aaatgctttg cacgtgcctg
     2041 ggcaacggag tcagctgcca agagacagct gtaacccaga cttacggtgg caactcaaat
     2101 ggagagccat gtgtcttacc attcacctac aatggcagga cgttctactc ctgcaccaca
     2161 gaagggcgac aggacggaca tctttggtgc agcacaactt cgaattatga gcaggaccag
     2221 aaatactctt tctgcacaga ccacactgtt ttggttcaga ctcgaggagg aaattccaat
     2281 ggtgccttgt gccacttccc cttcctatac aacaaccaca attacactga ttgcacttct
     2341 gagggcagaa gagacaacat gaagtggtgt gggaccacac agaactatga tgccgaccag
     2401 aagtttgggt tctgccccat ggctgcccac gaggaaatct gcacaaccaa tgaaggggtc
     2461 atgtaccgca ttggagatca gtgggataag cagcatgaca tgggtcacat gatgaggtgc
     2521 acgtgtgttg ggaatggtcg tggggaatgg acatgcattg cctactcgca gcttcgagat
     2581 cagtgcattg ttgatgacat cacttacaat gtgaacgaca cattccacaa gcgtcatgaa
     2641 gaggggcaca tgctgaactg tacatgcttc ggtcagggtc ggggcaggtg gaagtgtgat
     2701 cccgtcgacc aatgccagga ttcagagact gggacgtttt atcaaattgg agattcatgg
     2761 gagaagtatg tgcatggtgt cagataccag tgctactgct atggccgtgg cattggggag
     2821 tggcattgcc aacctttaca gacctatcca agctcaagtg gtcctgtcga agtatttatc
     2881 actgagactc cgagtcagcc caactcccac cccatccagt ggaatgcacc acagccatct
     2941 cacatttcca agtacattct caggtggaga cctaaaaatt ctgtaggccg ttggaaggaa
     3001 gctaccatac caggccactt aaattcctac accatcaaag gcctgaagcc tggtgtggta
     3061 tacgagggcc agctcatcag catccagcag tacggccacc aagaagtgac tcgctttgac
     3121 ttcaccacca ccagcaccag cacacctgtg accagcaaca ccgtgacagg agagacgact
     3181 cccttttctc ctcttgtggc cacttctgaa tctgtgaccg aaatcacagc cagtagcttt
     3241 gtggtctcct gggtctcagc ttccgacacc gtgtcgggat tccgggtgga atatgagctg
     3301 agtgaggagg gagatgagcc acagtacctg gatcttccaa gcacagccac ttctgtgaac
     3361 atccctgacc tgcttcctgg ccgaaaatac attgtaaatg tctatcagat atctgaggat
     3421 ggggagcaga gtttgatcct gtctacttca caaacaacag cgcctgatgc cccacctgac
     3481 ccgactgtgg accaagttga tgacacctca attgttgttc gctggagcag accccaggct
     3541 cccatcacag ggtacagaat agtctattcg ccatcagtag aaggtagcag cacagaactc
     3601 aaccttcctg aaactgcaaa ctccgtcacc ctcagtgact tgcaacctgg tgttcagtat
     3661 aacatcgcta tctatgctgt ggaagaaaat caagaaagta cacctgttgt cattcaacaa
     3721 gaaaccactg gcaccccacg ctcagataca gtgccctctc ccagggacct gcagtttgtg
     3781 gaagtgacag acgtgaaggt caccatcatg tggacaccgc ctgagagtgc agtgaccggc
     3841 taccgtgtgg atgtgatccc cgtcaacctg cctggcgagc acgggcagag gctgcccatc
     3901 agcaggaaca cctttgcaga agtcaccggg ctgtcccctg gggtcaccta ttacttcaaa
     3961 gtctttgcag tgagccatgg gagggagagc aagcctctga ctgctcaaca gacaaccaaa
     4021 ctggatgctc ccactaacct ccagtttgtc aatgaaactg attctactgt cctggtgaga
     4081 tggactccac ctcgggccca gataacagga taccgactga ccgtgggcct tacccgaaga
     4141 ggacagccca ggcagtacaa tgtgggtccc tctgtctcca agtacccact gaggaatctg
     4201 cagcctgcat ctgagtacac cgtatccctc gtggccataa agggcaacca agagagcccc
     4261 aaagccactg gagtctttac cacactgcag cctgggagct ctattccacc ttacaacacc
     4321 gaggtgactg agaccaccat tgtgatcaca tggacgcctg ctccaagaat tggttttaag
     4381 ctgggtgtac gaccaagcca gggaggagag gcaccacgag aagtgacttc agactcagga
     4441 agcatcgttg tgtccggctt gactccagga gtagaatacg tctacaccat ccaagtcctg
     4501 agagatggac aggaaagaga tgcgccaatt gtaaacaaag tggtgacacc attgtctcca
     4561 ccaacaaact tgcatctgga ggcaaaccct gacactggag tgctcacagt ctcctgggag
     4621 aggagcacca ccccagacat tactggttat agaattacca caacccctac aaacggccag
     4681 cagggaaatt ctttggaaga agtggtccat gctgatcaga gctcctgcac ttttgataac
     4741 ctgagtcccg gcctggagta caatgtcagt gtttacactg tcaaggatga caaggaaagt
     4801 gtccctatct ctgataccat catcccagct gttcctcctc ccactgacct gcgattcacc
     4861 aacattggtc cagacaccat gcgtgtcacc tgggctccac ccccatccat tgatttaacc
     4921 aacttcctgg tgcgttactc acctgtgaaa aatgaggaag atgttgcaga gttgtcaatt
     4981 tctccttcag acaatgcagt ggtcttaaca aatctcctgc ctggtacaga atatgtagtg
     5041 agtgtctcca gtgtctacga acaacatgag agcacacctc ttagaggaag acagaaaaca
     5101 ggtcttgatt ccccaactgg cattgacttt tctgatatta ctgccaactc ttttactgtg
     5161 cactggattg ctcctcgagc caccatcact ggctacagga tccgccatca tcccgagcac
     5221 ttcagtggga gacctcgaga agatcgggtg ccccactctc ggaattccat caccctcacc
     5281 aacctcactc caggcacaga gtatgtggtc agcatcgttg ctcttaatgg cagagaggaa
     5341 agtcccttat tgattggcca acaatcaaca gtttctgatg ttccgaggga cctggaagtt
     5401 gttgctgcga cccccaccag cctactgatc agctgggatg ctcctgctgt cacagtgaga
     5461 tattacagga tcacttacgg agaaacagga ggaaatagcc ctgtccagga gttcactgtg
     5521 cctgggagca agtctacagc taccatcagc ggccttaaac ctggagttga ttataccatc
     5581 actgtgtatg ctgtcactgg ccgtggagac agccccgcaa gcagcaagcc aatttccatt
     5641 aattaccgaa cagaaattga caaaccatcc cagatgcaag tgaccgatgt tcagggcaac
     5701 agcattagtg tcaagtggct gccttcaagt tcccctgtta ctggttacag agtaaccacc
     5761 actcccaaaa atggaccagg accaacaaaa actaaaactg caggtccaga tcaaacagaa
     5821 atgactattg aaggcttgca gcccacagtg gagtatgtgg ttagtgtcta tgctcagaat
     5881 ccaagcggag agagtcagcc tctggttcag actgcagtaa ccactattcc tgcaccaact
     5941 gacctgaagt tcactcaggt cacacccaca agcctgagcg cccagtggac accacccaat
     6001 gttcagctca ctggatatcg agtgcgggtg acccccaagg agaagaccgg accaatgaaa
     6061 gaaatcaacc ttgctcctga cagctcatcc gtggttgtat caggacttat ggtggccacc
     6121 aaatatgaag tgagtgtcta tgctcttaag gacactttga caagcagacc agctcagggt
     6181 gttgtcacca ctctggagaa tgtcagccca ccaagaaggg ctcgtgtgac agatgctact
     6241 gagaccacca tcaccattag ctggagaacc aagactgaga cgatcactgg cttccaagtt
     6301 gatgccgttc cagccaatgg ccagactcca atccagagaa ccatcaagcc agatgtcaga
     6361 agctacacca tcacaggttt acaaccaggc actgactaca agatctacct gtacaccttg
     6421 aatgacaatg ctcggagctc ccctgtggtc atcgacgcct ccactgccat tgatgcacca
     6481 tccaacctgc gtttcctggc caccacaccc aattccttgc tggtatcatg gcagccgcca
     6541 cgtgccagga ttaccggcta catcatcaag tatgagaagc ctgggtctcc tcccagagaa
     6601 gtggtccctc ggccccgccc tggtgtcaca gaggctacta ttactggcct ggaaccggga
     6661 accgaatata caatttatgt cattgccctg aagaataatc agaagagcga gcccctgatt
     6721 ggaaggaaaa agacagacga gcttccccaa ctggtaaccc ttccacaccc caatcttcat
     6781 ggaccagaga tcttggatgt tccttccaca gttcaaaaga cccctttcgt ctcccaccct
     6841 gggtatgaca ctggaaatgg tattcagctt cctggcactt ctggtcagca acccagtgtt
     6901 gggcaacaaa tgatctttga ggaacatggt tttaggcgga ccacaccgcc ctcatcggcc
     6961 acccccataa ggcataggcc aagaccatac ccgccgaatg taggtgagga aatccaaatt
     7021 ggtcacatcc ccagggaaga tgtagactat cacctgtacc cacacggtcc gggactcaat
     7081 ccaaatgcct ctacaggaca agaagctctc tctcagacaa ccatctcatg ggccccattc
     7141 caggacactt ctgagtacat catttcatgt catcctgttg gcactgatga agaaccctta
     7201 cagttcaggg ttcctggaac ttctaccagt gccactctga caggcctcac cagaggtgcc
     7261 acctacaaca tcatagtgga ggcactgaaa gaccagcaga ggcataaggt tcgggaagag
     7321 gttgttaccg tgggcaactc tgtcaacgaa ggcttgaacc aacctacgga tgactcgtgc
     7381 tttgacccct acacagtttc ccattatgcc gttggagatg agtgggaacg aatgtctgaa
     7441 tcaggcttta aactgttgtg ccagtgctta ggctttggaa gtggtcattt cagatgtgat
     7501 tcatctagat ggtgccatga caatggtgtg aactacaaga ttggagagaa gtgggaccgt
     7561 cagggagaaa atggccagat gatgagctgc acatgtcttg ggaacggaaa aggagaattc
     7621 aagtgtgacc ctcatgaggc aacgtgttat gatgatggga agacatacca cgtaggagaa
     7681 cagtggcaga aggaatatct cggtgccatt tgctcctgca catgctttgg aggccagcgg
     7741 ggctggcgct gtgacaactg ccgcagacct gggggtgaac ccactcccga aggcactact
     7801 ggccagtcct acaaccagta ttctcagaga taccatcaga gaacaaacac taatgttaat
     7861 tgcccaattg agtgcttcat gcctttagat gtacaggctg acagagaaga ttcccgagag
     7921 taaatcatct ttgcggccgc gtttaaacag agctcgatga gtttggacaa accacaacta
     7981 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     8041 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     8101 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtactt
     8161 aagaggggga gaccaaaggg cgagacgtta aggcctcacg tgacatgtga gcaaaaggcc
     8221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8401 tgccgcttac gggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgccg tctcagaaga
     8941 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     9001 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     9061 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     9121 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     9181 cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     9241 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     9301 tctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     9361 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     9421 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     9481 cgtcgagtac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     9541 cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     9601 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     9661 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     9721 caatcataat attattgaag catttatcag ggttcgtctc gtcccggtct cctcccatgc
     9781 atg
//
