LOCUS       sequence-343520-        6421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5-endoNA2 fusion protein for detection of polysialic 
            acid. The endoNA2 is a catalytically inactive form of endosialidase 
            from bacteriophage PK1A2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6421)
  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 6421)
  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: pV5endoNA2
FEATURES             Location/Qualifiers
     source          1..6421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..30
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(54..68,69..74,75..91,92..98)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    74..90
                     /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    106..122
                     /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             141..152
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             171..188
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      185..194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             198..239
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      3212..3306
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3350..4009
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3416..3435)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4074..4160
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4207..4229)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4315..4455
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4370..4389
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4470..4487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4641..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5400..6191,6192..6260))
                     /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     6023..6042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6261..6365)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag aaatcataaa
       61 aaatttattt gctttgtgag cggataacaa ttataataga ttcaattgtg agcggataac
      121 aatttcacac agaattcatt aaagaggaga aattaactat gagaggatct caccatcacc
      181 atcaccatgg cggccgcggt aagcctatcc ctaaccctct cctcggtctc gattctacga
      241 ctagtgggat cgcaaataac tatactgata gtaaaagtgc tgaaacattg gaaaatgcgc
      301 gtaaagaggc ttttgctcag tctaaagatg ttttgaatat ggcaaaagca cactcaaata
      361 gtgttgctag aacaacttta gaaactgctg aagaacatgc aaatagtgtt gccagaacaa
      421 ctttagaaac tgctgaagaa catgcaaata aaaaatcagc tgaggcgtta gcaagcgcta
      481 atgtgtatgc agacagcaag tcttctcaca cactaaaaac tgcaaatagc tataccgatg
      541 tgactgtaag taattcgact aagaaagcaa tccgtgaatc gaatcaatac acagatcata
      601 aattccgtca acttgacaac cggttagata agctgtacac ggatcaattc aagagtaaaa
      661 tagcaggtgc aatctggcgt aacttggatg acaagctcac cgaggttgta tcgcttaaag
      721 attttggagc caaaggtgat ggtaagacaa acgaccaaga tgcagtaaat gcagcgatgg
      781 cttcaggtaa gagaattgac ggtgctggtg ctacttacaa agtatcatct ttacctgata
      841 tggagcgatt ctataacacc cgcttcgtat gggaacgttt atcaggtcaa cctctttact
      901 atgtgagtaa aggttttatc aatggtgaac tctataaaat cacggataac ccttattaca
      961 atgcttggcc tcaagacaaa gcgtttgtat atgagaacgt gatatatgca ccttacatgg
     1021 gtagcgaccg tcatggtgtt agtcgtctgc atgtatcatg ggttaagtct ggtgacgatg
     1081 gtcaaacatg gtctacacca gagtggttaa ctgatctgca tccagattac cctacagtga
     1141 actatcattg tatgagcatg ggtgtatgtc gcaaccgtct gtttgccatg attgaaacac
     1201 gtactttagc caagaacgca ctaaccaatt gtgcattgtg ggatcgccct atgtctcgta
     1261 gtctgcatct tacgggtggt atcactaagg ctgcaaatca gcgatatgca acaatacatg
     1321 taccagatca cggactattc gtgggcgatt ttgttaactt ctctaattct gcggtaacag
     1381 gtgtatcagg tgatatgact gttgcaacgg taatagataa ggacaacttc acggttctta
     1441 cacctaacca gcagacttca gatttgaata acgctggaaa gaattggcac atgggtactt
     1501 ctttccataa gtctccgtgg cgtaagacag atcttggtct aatccctcat gtcacagagg
     1561 tgcatagctt tgctactatt gataacaatg gctttgttat gggctatcat caaggtgatg
     1621 tagctccacg agaagttggg cttttctact tccctgatgc tttcaatagc ccatctaatt
     1681 atgttcgtcg tcagatacca tctgagtatg aaccagatgc ggcagagcca tgcatcaagt
     1741 actatgacgg tgtattatac cttatcactc gtggtactcg tggcgaccga ctaggaagct
     1801 ctctgcatcg tagtagagat ataggtcaga cttgggagtc actaagattt ccacataatg
     1861 tgtatcatac tactttaccg tttgctaagg taggagatga ccttattatg tttggttcag
     1921 aacgtgcaga aaatgaatgg gaagcaggtg caccagatga tcgttacaag gcatcttatc
     1981 ctcgtacctt ctatgcacga ttgaatgtaa acaattggaa tgcagctgat attgaatggg
     2041 ttaacatcac agaccaaatc tatcagggtg acattgtgga ctctagtgta ggtgtaggtt
     2101 cggtagtagt taaagacagc tacatttact atatctttgg tggcgaaaac catttcaacc
     2161 caatgactta tggtgacaac agagataaag acccatttaa aggtcatgga caccctactg
     2221 atatatactg ctataagatg cagattgcaa atgacagtcg tgtatctcgt aagtttacat
     2281 atggtgcaac tcccggacag gctataccta ccttcatggg tactgatggt attcgtacta
     2341 ttcctgctcc acttcactta tcaggagagg tagttgccgc agatatgact gtcgaacatc
     2401 ttaccctgaa ggcaagtaca agtgctaata ttcgctcaga gatgctgatg gaaggtgaat
     2461 atgggttcat tggtaagact ataccaaagg ataacccaac tgcccaacgt gtcatcatct
     2521 ctggtggagg gggtacagca gcagataccg gtgcgcaaat taccttacat ggagcaggtt
     2581 ctagtacttc tagacgcgca gtatataatg ccaatgagca cttgttccaa agcggggcta
     2641 tcatgcccta caatgataac gtgtatcctg ctggcggccc tcgtaataga ttcactacta
     2701 tccatcttac aagtgaccct attattacgt ctgatgccac tcataagtat ggatcacaag
     2761 gtattgatga gagtgtacta aaggcatggg gtaaggttag cttcaagcag tacaagttga
     2821 ttggtgagat gtctcgtggt gtgcaccaca ctcactttgg tgtattggca caagacattg
     2881 tagctgcttt tgcatccgaa gggttagatg caatcgactt cggcattgtc tccttcgaag
     2941 aaggtcagtt tggtgttaga tatagtgaaa tcttaatctt ggaggctgct tatactcgtt
     3001 atcgtttaga taagttagag gagatgtatg ccactaataa aatcagttaa gcaagctgct
     3061 gtactccaga atacagaaga acttattcaa tcaggacgtg accctaagca ggcatacgcc
     3121 attgccaagg atgtacaagt aaagcggccg atccgcatgc gagctcggta ccccgggtcg
     3181 acctgcagcc aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
     3241 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     3301 gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     3361 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     3421 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     3481 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     3541 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     3601 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     3661 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     3721 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     3781 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     3841 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     3901 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     3961 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     4021 cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa
     4081 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     4141 ctctcctgag taggacaaat ccgccctcta gagctgcctc gcgcgtttcg gtgatgacgg
     4201 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4261 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     4321 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     4381 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4441 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6361 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     6421 t
//
