LOCUS       sequence-343523-        7546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DT386C-endoNA2 fusion protein for selective elimination of
            polysialic acid–containing cells. Truncated diphtheria toxin fused 
            to noncatalytic endosialidase via a caspase-3/7-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: pDTCendoNA2
FEATURES             Location/Qualifiers
     source          1..7546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..28
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(52..66,67..72,73..89,90..96)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    72..88
                     /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    104..120
                     /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             139..150
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             169..186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      183..192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             202..774
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     terminator      4335..4429
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4473..5132
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4539..4558)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5197..5283
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5330..5352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5438..5578
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5493..5512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5593..5610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5764..6352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5844..5863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6523..7314,7315..7383))
                     /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     7146..7165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7384..7488)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctataaaa ataggcgtat cacgaggccc tttcgtcttc acctcgagaa atcataaaaa
       61 atttatttgc tttgtgagcg gataacaatt ataatagatt caattgtgag cggataacaa
      121 tttcacacag aattcattaa agaggagaaa ttaactatga gaggatctca ccatcaccat
      181 caccatggcg gccgcggcgc tgatgatgtt gttgattctt ctaaatcttt tgtgatggaa
      241 aacttttctt cgtaccacgg gactaaacct ggttatgtag attccattca aaaaggtata
      301 caaaagccaa aatctggtac acaaggaaat tatgacgatg attggaaagg gttttatagt
      361 accgacaata aatacgacgc tgcgggatac tctgtagata atgaaaaccc gctctctgga
      421 aaagctggag gcgtggtcaa agtgacgtat ccaggactga cgaaggttct cgcactaaaa
      481 gtggataatg ccgaaactat taagaaagag ttaggtttaa gtctcactga accgttgatg
      541 gagcaagtcg gaacggaaga gtttatcaaa aggttcggtg atggtgcttc gcgtgtagtg
      601 ctcagccttc ccttcgctga ggggagttct agcgttgaat atattaataa ctgggaacag
      661 gcgaaagcgt taagcgtaga acttgagatt aattttgaaa cccgtggaaa acgtggccaa
      721 gatgcgatgt atgagtatat ggctcaagcc tgtgcaggaa atcgtgtcag gcgatcagta
      781 ggtagctcat tgtcatgcat aaatcttgat tgggatgtca taagggataa aactaagaca
      841 aagatagagt ctttgaaaga gcatggccct atcaaaaata aaatgagcga aagtcccaat
      901 aaaacagtat ctgaggaaaa agctaaacaa tacctagaag aatttcatca aacggcatta
      961 gagcatcctg aattgtcaga acttaaaacc gttactggga ccaatcctgt attcgctggg
     1021 gctaactatg cggcgtgggc agtaaacgtt gcgcaagtta tcgatagcga aacagctgat
     1081 aatttggaaa agacaactgc tgctctttcg atacttcctg gtatcggtag cgtaatgggc
     1141 attgcagacg gtgccgttca ccacaataca gaagagatag tggcacaatc aatagcttta
     1201 tcgtctttaa tggttgctca agctattcca ttggtaggag agctagttga tattggtttc
     1261 gctgcatata attttgtaga gagtattatc aatttatttc aagtagttca taattcgtat
     1321 aatcgtcccg cgtattctcc ggggcataaa gacgaagttg acactagtgg gatcgcaaat
     1381 aactatactg atagtaaaag tgctgaaaca ttggaaaatg cgcgtaaaga ggcttttgct
     1441 cagtctaaag atgttttgaa tatggcaaaa gcacactcaa atagtgttgc tagaacaact
     1501 ttagaaactg ctgaagaaca tgcaaatagt gttgccagaa caactttaga aactgctgaa
     1561 gaacatgcaa ataaaaaatc agctgaggcg ttagcaagcg ctaatgtgta tgcagacagc
     1621 aagtcttctc acacactaaa aactgcaaat agctataccg atgtgactgt aagtaattcg
     1681 actaagaaag caatccgtga atcgaatcaa tacacagatc ataaattccg tcaacttgac
     1741 aaccggttag ataagctgta cacggatcaa ttcaagagta aaatagcagg tgcaatctgg
     1801 cgtaacttgg atgacaagct caccgaggtt gtatcgctta aagattttgg agccaaaggt
     1861 gatggtaaga caaacgacca agatgcagta aatgcagcga tggcttcagg taagagaatt
     1921 gacggtgctg gtgctactta caaagtatca tctttacctg atatggagcg attctataac
     1981 acccgcttcg tatgggaacg tttatcaggt caacctcttt actatgtgag taaaggtttt
     2041 atcaatggtg aactctataa aatcacggat aacccttatt acaatgcttg gcctcaagac
     2101 aaagcgtttg tatatgagaa cgtgatatat gcaccttaca tgggtagcga ccgtcatggt
     2161 gttagtcgtc tgcatgtatc atgggttaag tctggtgacg atggtcaaac atggtctaca
     2221 ccagagtggt taactgatct gcatccagat taccctacag tgaactatca ttgtatgagc
     2281 atgggtgtat gtcgcaaccg tctgtttgcc atgattgaaa cacgtacttt agccaagaac
     2341 gcactaacca attgtgcatt gtgggatcgc cctatgtctc gtagtctgca tcttacgggt
     2401 ggtatcacta aggctgcaaa tcagcgatat gcaacaatac atgtaccaga tcacggacta
     2461 ttcgtgggcg attttgttaa cttctctaat tctgcggtaa caggtgtatc aggtgatatg
     2521 actgttgcaa cggtaataga taaggacaac ttcacggttc ttacacctaa ccagcagact
     2581 tcagatttga ataacgctgg aaagaattgg cacatgggta cttctttcca taagtctccg
     2641 tggcgtaaga cagatcttgg tctaatccct catgtcacag aggtgcatag ctttgctact
     2701 attgataaca atggctttgt tatgggctat catcaaggtg atgtagctcc acgagaagtt
     2761 gggcttttct acttccctga tgctttcaat agcccatcta attatgttcg tcgtcagata
     2821 ccatctgagt atgaaccaga tgcggcagag ccatgcatca agtactatga cggtgtatta
     2881 taccttatca ctcgtggtac tcgtggcgac cgactaggaa gctctctgca tcgtagtaga
     2941 gatataggtc agacttggga gtcactaaga tttccacata atgtgtatca tactacttta
     3001 ccgtttgcta aggtaggaga tgaccttatt atgtttggtt cagaacgtgc agaaaatgaa
     3061 tgggaagcag gtgcaccaga tgatcgttac aaggcatctt atcctcgtac cttctatgca
     3121 cgattgaatg taaacaattg gaatgcagct gatattgaat gggttaacat cacagaccaa
     3181 atctatcagg gtgacattgt ggactctagt gtaggtgtag gttcggtagt agttaaagac
     3241 agctacattt actatatctt tggtggcgaa aaccatttca acccaatgac ttatggtgac
     3301 aacagagata aagacccatt taaaggtcat ggacacccta ctgatatata ctgctataag
     3361 atgcagattg caaatgacag tcgtgtatct cgtaagttta catatggtgc aactcccgga
     3421 caggctatac ctaccttcat gggtactgat ggtattcgta ctattcctgc tccacttcac
     3481 ttatcaggag aggtagttgc cgcagatatg actgtcgaac atcttaccct gaaggcaagt
     3541 acaagtgcta atattcgctc agagatgctg atggaaggtg aatatgggtt cattggtaag
     3601 actataccaa aggataaccc aactgcccaa cgtgtcatca tctctggtgg agggggtaca
     3661 gcagcagata ccggtgcgca aattacctta catggagcag gttctagtac ttctagacgc
     3721 gcagtatata atgccaatga gcacttgttc caaagcgggg ctatcatgcc ctacaatgat
     3781 aacgtgtatc ctgctggcgg ccctcgtaat agattcacta ctatccatct tacaagtgac
     3841 cctattatta cgtctgatgc cactcataag tatggatcac aaggtattga tgagagtgta
     3901 ctaaaggcat ggggtaaggt tagcttcaag cagtacaagt tgattggtga gatgtctcgt
     3961 ggtgtgcacc acactcactt tggtgtattg gcacaagaca ttgtagctgc ttttgcatcc
     4021 gaagggttag atgcaatcga cttcggcatt gtctccttcg aagaaggtca gtttggtgtt
     4081 agatatagtg aaatcttaat cttggaggct gcttatactc gttatcgttt agataagtta
     4141 gaggagatgt atgccactaa taaaatcagt taagcaagct gctgtactcc agaatacaga
     4201 agaacttatt caatcaggac gtgaccctaa gcaggcatac gccattgcca aggatgtaca
     4261 agtaaagcgg ccgatccgca tgcgagctcg gtaccccggg tcgacctgca gccaagctta
     4321 attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     4381 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagct
     4441 tggcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     4501 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     4561 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     4621 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     4681 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     4741 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     4801 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     4861 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     4921 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     4981 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     5041 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     5101 agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt
     5161 aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg
     5221 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     5281 aatccgccct ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5341 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5401 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     5461 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     5521 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6181 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7201 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7261 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7321 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7501 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acatta
//
