LOCUS       sequence-343522-        7546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DT386F-endoNA2 fusion protein for selective elimination of
            polysialic acid–containing cells. Truncated diphtheria toxin fused 
            to noncatalytic endosialidase via a furin-cleavable linker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7546)
  AUTHORS   Lehti TA, Pajunen MI, Jokilammi A, Korja M, Lilie H, Vettenranta K, 
            Finne J
  TITLE     Design of a cytotoxic neuroblastoma-targeting agent using an enzyme 
            acting on polysialic acid fused to a toxin.
  JOURNAL   Mol Cancer Ther. 2021 Jul 26. pii: 1535-7163.MCT-20-1031. doi: 
            10.1158/1535-7163.MCT-20-1031.
  PUBMED    34315766
REFERENCE   2  (bases 1 to 7546)
  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: "Mol Cancer 
            Ther. 2021 Jul 26. pii: 1535-7163.MCT-20-1031. doi: 
            10.1158/1535-7163.MCT-20-1031."
COMMENT     Sequence Label: pDTFendoNA2
FEATURES             Location/Qualifiers
     source          1..7546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(58..72,73..78,79..95,96..102)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    78..94
                     /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)."
     protein_bind    110..126
                     /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)."
     RBS             145..156
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             175..192
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      189..198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             208..780
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     terminator      4341..4435
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4479..5138
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4545..4564)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5203..5289
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5336..5358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5444..5584
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5499..5518
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5599..5616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5770..6358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5850..5869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6529..7320,7321..7389))
                     /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     7152..7171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7390..7494)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagaaatca
       61 taaaaaattt atttgctttg tgagcggata acaattataa tagattcaat tgtgagcgga
      121 taacaatttc acacagaatt cattaaagag gagaaattaa ctatgagagg atctcaccat
      181 caccatcacc atggcggccg cggcgctgat gatgttgttg attcttctaa atcttttgtg
      241 atggaaaact tttcttcgta ccacgggact aaacctggtt atgtagattc cattcaaaaa
      301 ggtatacaaa agccaaaatc tggtacacaa ggaaattatg acgatgattg gaaagggttt
      361 tatagtaccg acaataaata cgacgctgcg ggatactctg tagataatga aaacccgctc
      421 tctggaaaag ctggaggcgt ggtcaaagtg acgtatccag gactgacgaa ggttctcgca
      481 ctaaaagtgg ataatgccga aactattaag aaagagttag gtttaagtct cactgaaccg
      541 ttgatggagc aagtcggaac ggaagagttt atcaaaaggt tcggtgatgg tgcttcgcgt
      601 gtagtgctca gccttccctt cgctgagggg agttctagcg ttgaatatat taataactgg
      661 gaacaggcga aagcgttaag cgtagaactt gagattaatt ttgaaacccg tggaaaacgt
      721 ggccaagatg cgatgtatga gtatatggct caagcctgtg caggaaatcg tgtcaggcga
      781 tcagtaggta gctcattgtc atgcataaat cttgattggg atgtcataag ggataaaact
      841 aagacaaaga tagagtcttt gaaagagcat ggccctatca aaaataaaat gagcgaaagt
      901 cccaataaaa cagtatctga ggaaaaagct aaacaatacc tagaagaatt tcatcaaacg
      961 gcattagagc atcctgaatt gtcagaactt aaaaccgtta ctgggaccaa tcctgtattc
     1021 gctggggcta actatgcggc gtgggcagta aacgttgcgc aagttatcga tagcgaaaca
     1081 gctgataatt tggaaaagac aactgctgct ctttcgatac ttcctggtat cggtagcgta
     1141 atgggcattg cagacggtgc cgttcaccac aatacagaag agatagtggc acaatcaata
     1201 gctttatcgt ctttaatggt tgctcaagct attccattgg taggagagct agttgatatt
     1261 ggtttcgctg catataattt tgtagagagt attatcaatt tatttcaagt agttcataat
     1321 tcgtataatc gtcccgcgta ttctccgggg cataaacgtg tcaggcgaac tagtgggatc
     1381 gcaaataact atactgatag taaaagtgct gaaacattgg aaaatgcgcg taaagaggct
     1441 tttgctcagt ctaaagatgt tttgaatatg gcaaaagcac actcaaatag tgttgctaga
     1501 acaactttag aaactgctga agaacatgca aatagtgttg ccagaacaac tttagaaact
     1561 gctgaagaac atgcaaataa aaaatcagct gaggcgttag caagcgctaa tgtgtatgca
     1621 gacagcaagt cttctcacac actaaaaact gcaaatagct ataccgatgt gactgtaagt
     1681 aattcgacta agaaagcaat ccgtgaatcg aatcaataca cagatcataa attccgtcaa
     1741 cttgacaacc ggttagataa gctgtacacg gatcaattca agagtaaaat agcaggtgca
     1801 atctggcgta acttggatga caagctcacc gaggttgtat cgcttaaaga ttttggagcc
     1861 aaaggtgatg gtaagacaaa cgaccaagat gcagtaaatg cagcgatggc ttcaggtaag
     1921 agaattgacg gtgctggtgc tacttacaaa gtatcatctt tacctgatat ggagcgattc
     1981 tataacaccc gcttcgtatg ggaacgttta tcaggtcaac ctctttacta tgtgagtaaa
     2041 ggttttatca atggtgaact ctataaaatc acggataacc cttattacaa tgcttggcct
     2101 caagacaaag cgtttgtata tgagaacgtg atatatgcac cttacatggg tagcgaccgt
     2161 catggtgtta gtcgtctgca tgtatcatgg gttaagtctg gtgacgatgg tcaaacatgg
     2221 tctacaccag agtggttaac tgatctgcat ccagattacc ctacagtgaa ctatcattgt
     2281 atgagcatgg gtgtatgtcg caaccgtctg tttgccatga ttgaaacacg tactttagcc
     2341 aagaacgcac taaccaattg tgcattgtgg gatcgcccta tgtctcgtag tctgcatctt
     2401 acgggtggta tcactaaggc tgcaaatcag cgatatgcaa caatacatgt accagatcac
     2461 ggactattcg tgggcgattt tgttaacttc tctaattctg cggtaacagg tgtatcaggt
     2521 gatatgactg ttgcaacggt aatagataag gacaacttca cggttcttac acctaaccag
     2581 cagacttcag atttgaataa cgctggaaag aattggcaca tgggtacttc tttccataag
     2641 tctccgtggc gtaagacaga tcttggtcta atccctcatg tcacagaggt gcatagcttt
     2701 gctactattg ataacaatgg ctttgttatg ggctatcatc aaggtgatgt agctccacga
     2761 gaagttgggc ttttctactt ccctgatgct ttcaatagcc catctaatta tgttcgtcgt
     2821 cagataccat ctgagtatga accagatgcg gcagagccat gcatcaagta ctatgacggt
     2881 gtattatacc ttatcactcg tggtactcgt ggcgaccgac taggaagctc tctgcatcgt
     2941 agtagagata taggtcagac ttgggagtca ctaagatttc cacataatgt gtatcatact
     3001 actttaccgt ttgctaaggt aggagatgac cttattatgt ttggttcaga acgtgcagaa
     3061 aatgaatggg aagcaggtgc accagatgat cgttacaagg catcttatcc tcgtaccttc
     3121 tatgcacgat tgaatgtaaa caattggaat gcagctgata ttgaatgggt taacatcaca
     3181 gaccaaatct atcagggtga cattgtggac tctagtgtag gtgtaggttc ggtagtagtt
     3241 aaagacagct acatttacta tatctttggt ggcgaaaacc atttcaaccc aatgacttat
     3301 ggtgacaaca gagataaaga cccatttaaa ggtcatggac accctactga tatatactgc
     3361 tataagatgc agattgcaaa tgacagtcgt gtatctcgta agtttacata tggtgcaact
     3421 cccggacagg ctatacctac cttcatgggt actgatggta ttcgtactat tcctgctcca
     3481 cttcacttat caggagaggt agttgccgca gatatgactg tcgaacatct taccctgaag
     3541 gcaagtacaa gtgctaatat tcgctcagag atgctgatgg aaggtgaata tgggttcatt
     3601 ggtaagacta taccaaagga taacccaact gcccaacgtg tcatcatctc tggtggaggg
     3661 ggtacagcag cagataccgg tgcgcaaatt accttacatg gagcaggttc tagtacttct
     3721 agacgcgcag tatataatgc caatgagcac ttgttccaaa gcggggctat catgccctac
     3781 aatgataacg tgtatcctgc tggcggccct cgtaatagat tcactactat ccatcttaca
     3841 agtgacccta ttattacgtc tgatgccact cataagtatg gatcacaagg tattgatgag
     3901 agtgtactaa aggcatgggg taaggttagc ttcaagcagt acaagttgat tggtgagatg
     3961 tctcgtggtg tgcaccacac tcactttggt gtattggcac aagacattgt agctgctttt
     4021 gcatccgaag ggttagatgc aatcgacttc ggcattgtct ccttcgaaga aggtcagttt
     4081 ggtgttagat atagtgaaat cttaatcttg gaggctgctt atactcgtta tcgtttagat
     4141 aagttagagg agatgtatgc cactaataaa atcagttaag caagctgctg tactccagaa
     4201 tacagaagaa cttattcaat caggacgtga ccctaagcag gcatacgcca ttgccaagga
     4261 tgtacaagta aagcggccga tccgcatgcg agctcggtac cccgggtcga cctgcagcca
     4321 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     4381 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     4441 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     4501 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     4561 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     4621 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     4681 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     4741 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     4801 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     4861 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     4921 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     4981 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     5041 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     5101 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     5161 gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct
     5221 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5281 aggacaaatc cgccctctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5341 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5401 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     5461 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     5521 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5581 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6181 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7201 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7381 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7441 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7501 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatg
//
