LOCUS       sequence-343066-        7082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-endoNA fusion protein for degradation of polysialic 
            acid. The endoNA is a catalytically active endosialidase from 
            bacteriophage PK1A..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  AUTHORS   Jokilammi A, Ollikka P, Korja M, Jakobsson E, Loimaranta V, Haataja 
            S, Hirvonen H, Finne J
  TITLE     Construction of antibody mimics from a noncatalytic enzyme-detection
            of polysialic acid.
  JOURNAL   J Immunol Methods. 2004 Dec;295(1-2):149-60. doi: 
            10.1016/j.jim.2004.10.006. Epub 2004 Nov 14.
  PUBMED    15627620
REFERENCE   2  (bases 1 to 7082)
  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: "J Immunol 
            Methods."; date: "2004-12"; volume: "295(1-2):149-60. doi"; pages: "
            10.1016/j.jim.2004.10.006. Epub 2004 Nov 14"
COMMENT     Sequence Label: pFEndoNA
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(203..222)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     550..571
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3557..3651
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3695..4354
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3761..3780)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4419..4505
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4552..4574)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4660..4800
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4715..4734
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4815..4832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4986..5574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5066..5085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5745..6536,6537..6605))
                     /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     6368..6387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6606..6710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6778..6796
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6820..6834,6835..6840,6841..6857,6858..6864)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    6840..6856
                     /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    6872..6888
                     /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             6907..6918
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6937..6954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      6973..6982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6979..6981,6982..6984,6985..7082,1..610)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(6979..6981,6982..6984,6985..7082,1..610)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(7024..7045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
       61 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      121 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      181 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      241 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      301 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      361 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      421 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      481 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      541 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      601 cgagctgtac gggatcgcaa ataactatac tgatagtaaa agtgctgaaa cattggaaaa
      661 tgcgcgtaaa gaggcttttg ctcagtctaa agatgttttg aatatggcaa aagcacactc
      721 aaatagtgtt gctagaacaa ctttagaaac tgctgaagaa catgcaaata gtgttgccag
      781 aacaacttta gaaactgctg aagaacatgc aaataaaaaa tcagctgagg cgttagcaag
      841 cgctaatgtg tatgcagaca gcaagtcttc tcacacacta aaaactgcaa atagctatac
      901 cgatgtgact gtaagtaatt cgactaagaa agcaatccgt gaatcgaatc aatacacaga
      961 tcataaattc cgtcaacttg acaaccggtt agataagctg tacacggatc aattcaagag
     1021 taaaatagca ggtgcaatct ggcgtaactt ggatgacaag ctcaccgagg ttgtatcgct
     1081 taaagatttt ggagccaaag gtgatggtaa gacaaacgac caagatgcag taaatgcagc
     1141 gatggcttca ggtaagagaa ttgacggtgc tggtgctact tacaaagtat catctttacc
     1201 tgatatggag cgattctata acacccgctt cgtatgggaa cgtttatcag gtcaacctct
     1261 ttactatgtg agtaaaggtt ttatcaatgg tgaactctat aaaatcacgg ataaccctta
     1321 ttacaatgct tggcctcaag acaaagcgtt tgtatatgag aacgtgatat atgcacctta
     1381 catgggtagc gaccgtcatg gtgttagtcg tctgcatgta tcatgggtta agtctggtga
     1441 cgatggtcaa acatggtcta caccagagtg gttaactgat ctgcatccag attaccctac
     1501 agtgaactat cattgtatga gcatgggtgt atgtcgcaac cgtctgtttg ccatgattga
     1561 aacacgtact ttagccaaga acgcactaac caattgtgca ttgtgggatc gccctatgtc
     1621 tcgtagtctg catcttacgg gtggtatcac taaggctgca aatcagcgat atgcaacaat
     1681 acatgtacca gatcacggac tattcgtggg cgattttgtt aacttctcta attctgcggt
     1741 aacaggtgta tcaggtgata tgactgttgc aacggtaata gataaggaca acttcacggt
     1801 tcttacacct aaccagcaga cttcagattt gaataacgct ggaaagaatt ggcacatggg
     1861 tacttctttc cataagtctc cgtggcgtaa gacagatctt ggtctaatcc ctcatgtcac
     1921 agaggtgcat agctttgcta ctattgataa caatggcttt gttatgggct atcatcaagg
     1981 tgatgtagct ccacgagaag ttgggctttt ctacttccct gatgctttca atagcccatc
     2041 taattatgtt cgtcgtcaga taccatctga gtatgaacca gatgcggcag agccatgcat
     2101 caagtactat gacggtgtat tataccttat cactcgtggt actcgtggcg accgactagg
     2161 aagctctctg catcgtagta gagatatagg tcagacttgg gagtcactaa gatttccaca
     2221 taatgtgcat catactactt taccgtttgc taaggtagga gatgacctta ttatgtttgg
     2281 ttcagaacgt gcagaaaatg aatgggaagc aggtgcacca gatgatcgtt acaaggcatc
     2341 ttatcctcgt accttctatg cacgattgaa tgtaaacaat tggaatgcag ctgatattga
     2401 atgggttaac atcacagacc aaatctatca gggtgacatt gtgaactcta gtgtaggtgt
     2461 aggttcggta gtagttaaag acagctacat ttactatatc tttggtggcg aaaaccattt
     2521 caacccaatg acttatggtg acaacagaga taaagaccca tttaaaggtc atggacaccc
     2581 tactgatata tactgctata agatgcagat tgcaaatgac agtcgtgtat ctcgtaagtt
     2641 tacatatggt gcaactcccg gacaggctat acctaccttc atgggtactg atggtattcg
     2701 tactattcct gctccacttc acttatcagg agaggtagtt gccgcagata tgactgtcga
     2761 acatcttacc ctgaaggcaa gtacaagtgc taatattcgc tcagagatgc tgatggaagg
     2821 tgaatatggg ttcattggta agactatacc aaaggataac ccaactgccc aacgtgtcat
     2881 catctctggt ggagggggta cagcagcaga taccggtgcg caaattacct tacatggagc
     2941 aggttctagt acttctagac gcgcagtata taatgccaat gagcacttgt tccaaagcgg
     3001 ggctatcatg ccctacaatg ataacgtgta tcctgctggc ggccctcgta atagattcac
     3061 tactatccat cttacaagtg accctattat tacgtctgat gccactcata agtatggatc
     3121 acaaggtatt gatgagagtg tactaaaggc atggggtaag gttagcttca agcagtacaa
     3181 gttgattggt gagatgtctc gtggtgtgca ccacactcac tttggtgtat tggcacaaga
     3241 cattgtagct gcttttgcat ccgaagggtt agatgcaatc gacttcggca ttgtctcctt
     3301 cgaagaaggt cagtttggtg ttagatatag tgaaatctta atcttggagg ctgcttatac
     3361 tcgttatcgt ttagataagt tagaggagat gtatgccact aataaaatca gttaagcaag
     3421 ctgctgtact ccagaataca gaagaactta ttcaatcagg acgtgaccct aagcaggcat
     3481 acgccattgc caaggatccg catgcgagct cggtaccccg ggtcgacctg cagccaagct
     3541 taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     3601 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag
     3661 cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     3721 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     3781 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     3841 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     3901 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     3961 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     4021 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     4081 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     4141 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     4201 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     4261 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     4321 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     4381 ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt
     4441 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     4501 caaatccgcc ctctagagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4561 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4621 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     4681 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4741 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4801 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4861 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5101 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5401 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5701 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5761 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5821 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5881 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5941 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6001 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6061 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6601 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6721 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6781 aggcgtatca cgaggccctt tcgtcttcac ctcgagaaat cataaaaaat ttatttgctt
     6841 tgtgagcgga taacaattat aatagattca attgtgagcg gataacaatt tcacacagaa
     6901 ttcattaaag aggagaaatt aactatgaga ggatctcacc atcaccatca ccatgatccc
     6961 cgggtaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     7021 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     7081 ga
//
