LOCUS       sequence-343513-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DsRed-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 7051)
  AUTHORS   Lehti TA, Pajunen MI, Skog MS, Finne J
  TITLE     Internalization of a polysialic acid-binding Escherichia coli 
            bacteriophage into eukaryotic neuroblastoma cells.
  JOURNAL   Nat Commun. 2017 Dec 4;8(1):1915. doi: 10.1038/s41467-017-02057-3.
  PUBMED    29203765
REFERENCE   2  (bases 1 to 7051)
  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: "Nat 
            Commun."; date: "2017-12-4"; volume: "8(1):1915. doi"; pages: " 
            10.1038/s41467-017-02057-3"
COMMENT     Sequence Label: pDsEndoNA2
FEATURES             Location/Qualifiers
     source          1..7051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..57
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(81..95,96..101,102..118,119..125)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    101..117
                     /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    133..149
                     /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             168..179
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             198..215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             225..896
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="DNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKL
                     QVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVV
                     EVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHA
                     LKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSG
                     SQ"
     primer_bind     620..639
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3869..3963
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4007..4666
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4073..4092)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4731..4817
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4864..4886)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4972..5112
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5027..5046
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5127..5144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5298..5886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5378..5397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6057..6848,6849..6917))
                     /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     6680..6699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6918..7022)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
       61 ttcgtcttca cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta
      121 taatagattc aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat
      181 taactatgag aggatctcac catcaccatc accatggcgg ccgcgacaac accgaggacg
      241 tcatcaagga gttcatgcag ttcaaggtgc gcatggaggg ctccgtgaac ggccactact
      301 tcgagatcga gggcgagggc gagggcaagc cctacgaggg cacccagacc gccaagctgc
      361 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt
      421 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacatg aagctgtcct
      481 tccccgaggg cttcacctgg gagcgctcca tgaacttcga ggacggcggc gtggtggagg
      541 tgcagcagga ctcctccctg caggacggca ccttcatcta caaggtgaag ttcaagggcg
      601 tgaacttccc cgccgacggc cccgtaatgc agaagaagac tgccggctgg gagccctcca
      661 ccgagaagct gtacccccag gacggcgtgc tgaagggcga gatctcccac gccctgaagc
      721 tgaaggacgg cggccactac acctgcgact tcaagaccgt gtacaaggcc aagaagcccg
      781 tgcagctgcc cggcaaccac tacgtggact ccaagctgga catcaccaac cacaacgagg
      841 actacaccgt ggtggagcag tacgagcacg ccgaggcccg ccactccggc tcccagacta
      901 gtgggatcgc aaataactat actgatagta aaagtgctga aacattggaa aatgcgcgta
      961 aagaggcttt tgctcagtct aaagatgttt tgaatatggc aaaagcacac tcaaatagtg
     1021 ttgctagaac aactttagaa actgctgaag aacatgcaaa tagtgttgcc agaacaactt
     1081 tagaaactgc tgaagaacat gcaaataaaa aatcagctga ggcgttagca agcgctaatg
     1141 tgtatgcaga cagcaagtct tctcacacac taaaaactgc aaatagctat accgatgtga
     1201 ctgtaagtaa ttcgactaag aaagcaatcc gtgaatcgaa tcaatacaca gatcataaat
     1261 tccgtcaact tgacaaccgg ttagataagc tgtacacgga tcaattcaag agtaaaatag
     1321 caggtgcaat ctggcgtaac ttggatgaca agctcaccga ggttgtatcg cttaaagatt
     1381 ttggagccaa aggtgatggt aagacaaacg accaagatgc agtaaatgca gcgatggctt
     1441 caggtaagag aattgacggt gctggtgcta cttacaaagt atcatcttta cctgatatgg
     1501 agcgattcta taacacccgc ttcgtatggg aacgtttatc aggtcaacct ctttactatg
     1561 tgagtaaagg ttttatcaat ggtgaactct ataaaatcac ggataaccct tattacaatg
     1621 cttggcctca agacaaagcg tttgtatatg agaacgtgat atatgcacct tacatgggta
     1681 gcgaccgtca tggtgttagt cgtctgcatg tatcatgggt taagtctggt gacgatggtc
     1741 aaacatggtc tacaccagag tggttaactg atctgcatcc agattaccct acagtgaact
     1801 atcattgtat gagcatgggt gtatgtcgca accgtctgtt tgccatgatt gaaacacgta
     1861 ctttagccaa gaacgcacta accaattgtg cattgtggga tcgccctatg tctcgtagtc
     1921 tgcatcttac gggtggtatc actaaggctg caaatcagcg atatgcaaca atacatgtac
     1981 cagatcacgg actattcgtg ggcgattttg ttaacttctc taattctgcg gtaacaggtg
     2041 tatcaggtga tatgactgtt gcaacggtaa tagataagga caacttcacg gttcttacac
     2101 ctaaccagca gacttcagat ttgaataacg ctggaaagaa ttggcacatg ggtacttctt
     2161 tccataagtc tccgtggcgt aagacagatc ttggtctaat ccctcatgtc acagaggtgc
     2221 atagctttgc tactattgat aacaatggct ttgttatggg ctatcatcaa ggtgatgtag
     2281 ctccacgaga agttgggctt ttctacttcc ctgatgcttt caatagccca tctaattatg
     2341 ttcgtcgtca gataccatct gagtatgaac cagatgcggc agagccatgc atcaagtact
     2401 atgacggtgt attatacctt atcactcgtg gtactcgtgg cgaccgacta ggaagctctc
     2461 tgcatcgtag tagagatata ggtcagactt gggagtcact aagatttcca cataatgtgt
     2521 atcatactac tttaccgttt gctaaggtag gagatgacct tattatgttt ggttcagaac
     2581 gtgcagaaaa tgaatgggaa gcaggtgcac cagatgatcg ttacaaggca tcttatcctc
     2641 gtaccttcta tgcacgattg aatgtaaaca attggaatgc agctgatatt gaatgggtta
     2701 acatcacaga ccaaatctat cagggtgaca ttgtggactc tagtgtaggt gtaggttcgg
     2761 tagtagttaa agacagctac atttactata tctttggtgg cgaaaaccat ttcaacccaa
     2821 tgacttatgg tgacaacaga gataaagacc catttaaagg tcatggacac cctactgata
     2881 tatactgcta taagatgcag attgcaaatg acagtcgtgt atctcgtaag tttacatatg
     2941 gtgcaactcc cggacaggct atacctacct tcatgggtac tgatggtatt cgtactattc
     3001 ctgctccact tcacttatca ggagaggtag ttgccgcaga tatgactgtc gaacatctta
     3061 ccctgaaggc aagtacaagt gctaatattc gctcagagat gctgatggaa ggtgaatatg
     3121 ggttcattgg taagactata ccaaaggata acccaactgc ccaacgtgtc atcatctctg
     3181 gtggaggggg tacagcagca gataccggtg cgcaaattac cttacatgga gcaggttcta
     3241 gtacttctag acgcgcagta tataatgcca atgagcactt gttccaaagc ggggctatca
     3301 tgccctacaa tgataacgtg tatcctgctg gcggccctcg taatagattc actactatcc
     3361 atcttacaag tgaccctatt attacgtctg atgccactca taagtatgga tcacaaggta
     3421 ttgatgagag tgtactaaag gcatggggta aggttagctt caagcagtac aagttgattg
     3481 gtgagatgtc tcgtggtgtg caccacactc actttggtgt attggcacaa gacattgtag
     3541 ctgcttttgc atccgaaggg ttagatgcaa tcgacttcgg cattgtctcc ttcgaagaag
     3601 gtcagtttgg tgttagatat agtgaaatct taatcttgga ggctgcttat actcgttatc
     3661 gtttagataa gttagaggag atgtatgcca ctaataaaat cagttaagca agctgctgta
     3721 ctccagaata cagaagaact tattcaatca ggacgtgacc ctaagcaggc atacgccatt
     3781 gccaaggatg tacaagtaaa gcggccgatc cgcatgcgag ctcggtaccc cgggtcgacc
     3841 tgcagccaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
     3901 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     3961 aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     4021 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     4081 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     4141 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     4201 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
     4261 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     4321 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     4381 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     4441 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     4501 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     4561 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     4621 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     4681 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
     4741 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4801 tcctgagtag gacaaatccg ccctctagag ctgcctcgcg cgtttcggtg atgacggtga
     4861 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4921 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4981 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     5041 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5101 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5701 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6661 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7021 cgcacatttc cccgaaaagt gccacctgac g
//
