LOCUS       sequence-251099-       10053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enables eukaryotic expression of human EDA (or EIIIA) containing 
            fibronectin based on plasma fibronectin sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10053)
  AUTHORS   Rossnagl S, Altrock E, Sens C, Kraft S, Rau K, Milsom MD, Giese T, 
            Samstag Y, Nakchbandi IA
  TITLE     EDA-Fibronectin Originating from Osteoblasts Inhibits the Immune 
            Response against Cancer.
  JOURNAL   PLoS Biol. 2016 Sep 21;14(9):e1002562. doi: 
            10.1371/journal.pbio.1002562. eCollection 2016 Sep.
  PUBMED    27653627
REFERENCE   2  (bases 1 to 10053)
  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: "PLoS 
            Biol."; date: "2016-09-21"; volume: "14(9):e1002562. doi"; pages: " 
            10.1371/journal.pbio.1002562. eCollection 2016 Sep"
COMMENT     Sequence Label: Fibronectin-human-EDA in pMAX
FEATURES             Location/Qualifiers
     source          1..10053
                     /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(8226..8245)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    8255..8336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     8280..8299
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8534..9122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8614..8633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9203..9997)
                     /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(9203..9997)
                     /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(9314..9333)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9924..9943
                     /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 cagaccacat 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 aaactcctac 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 aacatcacta 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 gggaatagcc ctgtccagga gttcactgtg
     5521 cctgggagca agtctacagc taccatcagc ggccttaaac ctggagttga ttataccatc
     5581 actgtgtatg ctgtcactgg ccgtggagac agccccgcaa gcagcaagcc aatttccatt
     5641 aattaccgaa cagaaattga caaaccatcc cagatgcaag tgaccgatgt tcaggacaac
     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 ccaacattga tcgccctaaa
     5941 ggactggcat tcactgatgt ggatgtcgat tccatcaaaa ttgcttggga aagcccacag
     6001 gggcaagttt ccaggtacag ggtgacctac tcgagccctg aggatggaat ccatgagcta
     6061 ttccctgcac ctgatggtga agaagacact gcagagctgc aaggcctcag accgggttct
     6121 gagtacacag tcagtgtggt tgccttgcac gatgatatgg agagccagcc cctgattgga
     6181 acccagtcca cagctattcc tgcaccaact gacctgaagt tcactcaggt cacacccaca
     6241 agcctgagcg cccagtggac accacccaat gttcagctca ctggatatcg agtgcgggtg
     6301 acccccaagg agaagaccgg accaatgaaa gaaatcaacc ttgctcctga cagctcatcc
     6361 gtggttgtat caggacttac ggtggccacc aaatatgaag tgagtgtcta tgctcttaag
     6421 gacactttga caagcagacc agctcaggga gttgtcacca ctctggagaa tgtcagccca
     6481 ccaagaaggg ctcgtgtgac agatgctact gagaccacca tcaccattag ctggagaacc
     6541 aagactgaga cgatcactgg cttccaagtt gatgccgttc cagccaatgg ccagactcca
     6601 atccagagaa ccatcaagcc agatgtcaga agctacacca tcacaggttt acaaccaggc
     6661 actgactaca agatctacct gtacaccttg aatgacaatg ctcggagctc ccctgtggtc
     6721 atcgacgcct ccactgccat tgatgcacca tccaacctgc gtttcctggc caccacaccc
     6781 aattccttgc tggtatcatg gcagccgcca cgtgccagga ttaccggcta catcatcaag
     6841 tatgagaagc ctgggtctcc tcccagagaa gtggtccctc ggccccgccc tggtgtcaca
     6901 gaggctacta ttactggcct ggaaccggga accgaatata caatttatgt cattgccctg
     6961 aagaataatc agaagagcga gcccctgatt ggaaggaaaa agacagacga gcttccccaa
     7021 ctggtaaccc ttccacaccc caatcttcat ggaccagaga tcttggatgt tccttccaca
     7081 gttcaaaaga cccctttcgt cacccaccct gggtatgaca ctggaaatgg tattcagctt
     7141 cctggcactt ctggtcagca acccagtgtt gggcaacaaa tgatctttga ggaacatggt
     7201 tttaggcgga ccacaccgcc cacaacggcc acccccataa ggcataggcc aagaccatac
     7261 ccgccgaatg taggtgagga aatccaaatt ggtcacatcc ccagggaaga tgtagactat
     7321 cacctgtacc cacacggtcc gggactcaat ccaaatgcct ctacaggaca agaagctctc
     7381 tctcagacaa ccatctcatg ggccccattc caggacactt ctgagtacat catttcatgt
     7441 catcctgttg gcactgatga agaaccctta cagttcaggg ttcctggaac ttctaccagt
     7501 gccactctga caggcctcac cagaggtgcc acctacaaca tcatagtgga ggcactgaaa
     7561 gaccagcaga ggcataaggt tcgggaagag gttgttaccg tgggcaactc tgtcaacgaa
     7621 ggcttgaacc aacctacgga tgactcgtgc tttgacccct acacagtttc ccattatgcc
     7681 gttggagatg agtgggaacg aatgtctgaa tcaggcttta aactgttgtg ccagtgctta
     7741 ggctttggaa gtggtcattt cagatgtgat tcatctagat ggtgccatga caatggtgtg
     7801 aactacaaga ttggagagaa gtgggaccgt cagggagaaa atggccagat gatgagctgc
     7861 acatgtcttg ggaacggaaa aggagaattc aagtgtgacc ctcatgaggc aacgtgttat
     7921 gatgatggga agacatacca cgtaggagaa cagtggcaga aggaatatct cggtgccatt
     7981 tgctcctgca catgctttgg aggccagcgg ggctggcgct gtgacaactg ccgcagacct
     8041 gggggtgaac ccactcccga aggcactact ggccagtcct acaaccagta ttctcagaga
     8101 taccatcaga gaacaaacac taatgttaat tgcccaattg agtgcttcat gcctttagat
     8161 gtacaggctg acagagaaga ttcccgagag taaatcatct ttgcggccgc gtttaaacag
     8221 agctcgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     8281 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     8341 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     8401 gcaagtaaaa cctctacaaa tgtggtactt aagaggggga gaccaaaggg cgagacgtta
     8461 aggcctcacg tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac gggatacctg tccgcctttc
     8701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8941 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     9001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9181 aagggatttt ggtcatgccg tctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     9241 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     9301 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     9361 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     9421 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     9481 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     9541 caagaccggc ttccatccga gtacgtgctc tctcgatgcg atgtttcgct tggtggtcga
     9601 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     9661 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     9721 gcagccagtc ccttcccgct tcagtgacaa cgtcgagtac agctgcgcaa ggaacgcccg
     9781 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     9841 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     9901 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     9961 ccggagaacc tgcgtgcaat ccatcttgtt caatcataat attattgaag catttatcag
    10021 ggttcgtctc gtcccggtct cctcccatgc atg
//
