LOCUS       Exported                7102 bp DNA     circular SYN 04-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7102)
  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 joulu;295:149-60
  PUBMED    15627620
REFERENCE   2  (bases 1 to 7102)
  AUTHORS   Finne lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.jim.2004.10.006"; journalName: "J Immunol Methods"; date:
            "2004-12"; volume: "295"; pages: "149-60"
COMMENT     host strain M15 pREP4
FEATURES             Location/Qualifiers
     source          1..7102
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..117
                     /codon_start=1
                     /label=start codon
                     /translation="M"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      163..172
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             169..882
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)
                     >gi|1674491|gb|U76561.1|CVU76561 Cloning vector pEGFP, 
                     complete sequence "
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             883..1272
                     /codon_start=1
                     /product=">gi|48606|emb|X13882.1| Y.enterocolitica yadA 
                     gene for YadA "
                     /label=yadA stalk
                     /translation="GIANNYTDSKSAETLENARKEAFAQSKDVLNMAKAHSNSVARTTL
                     ETAEEHANSVARTTLETAEEHANKKSAEALASANVYADSKSSHTLKTANSYTDVTVSNS
                     TKKAIRESNQYTDHKFRQLDNRLDKL"
     CDS             1285..3687
                     /codon_start=1
                     /product=">ATS93359.1 tail spike protein endosialidase 
                     [Escherichia phage mutPK1A2]
                     >MG004687.1:41538-43973 Escherichia phage mutPK1A2, 
                     complete genome"
                     /label=endoNA2
                     /translation="FKSKIAGAIWRNLDDKLTEVVSLKDFGAKGDGKTNDQDAVNAAMA
                     SGKRIDGAGATYKVSSLPDMERFYNTRFVWERLSGQPLYYVSKGFINGELYKITDNPYY
                     NAWPQDKAFVYENVIYAPYMGSDRHGVSRLHVSWVKSGDDGQTWSTPEWLTDLHPDYPT
                     VNYHCMSMGVCRNRLFAMIETRTLAKNALTNCALWDRPMSRSLHLTGGITKAANQRYAT
                     IHVPDHGLFVGDFVNFSNSAVTGVSGDMTVATVIDKDNFTVLTPNQQTSDLNNAGKNWH
                     MGTSFHKSPWRKTDLGLIPHVTEVHSFATIDNNGFVMGYHQGDVAPREVGLFYFPDAFN
                     SPSNYVRRQIPSEYEPDAAEPCIKYYDGVLYLITRGTRGDRLGSSLHRSRDIGQTWESL
                     RFPHNVYHTTLPFAKVGDDLIMFGSERAENEWEAGAPDDRYKASYPRTFYARLNVNNWN
                     AADIEWVNITDQIYQGDIVDSSVGVGSVVVKDSYIYYIFGGENHFNPMTYGDNRDKDPF
                     KGHGHPTDIYCYKMQIANDSRVSRKFTYGATPGQAIPTFMGTDGIRTIPAPLHLSGEVV
                     AADMTVEHLTLKASTSANIRSEMLMEREYGFIGKTIPKDNPTAQRVIISGGGGTAADTG
                     AQITLHGAGSSTSRRAVYNANEHLFQSGAIMPYNDNVYPAGGPRNRFTTIHLTSDPIIT
                     SDATHKYGSQGIDESVLKAWGKVSFKQYKLIGEMSRGVHHTHFGVLAQDIVAAFASEGL
                     DAIDFGIVSFEEGQFGVRYSEILILEAAYTRYRLDKLEEMYATNKIS"
     misc_feature    2500..2502
                     /label=H417Y
                     /note="CAT -> TAT (H417->Y) "
     misc_feature    2716..2718
                     /label=N489D
                     /note="AAC -> GAC (N489->D) "
     terminator      3849..3943
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3987..4646
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      4711..4797
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4952..5092
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5278..5866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6037..6897)
                     /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"
     promoter        complement(6898..7002)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatgatccc cgggtaccgg tcgccaccat ggtgagcaag
      181 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      241 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      301 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      361 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      421 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      481 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      541 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      601 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
      661 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      721 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
      781 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      841 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acgggatcgc aaataactat
      901 actgatagta aaagtgctga aacattggaa aatgcgcgta aagaggcttt tgctcagtct
      961 aaagatgttt tgaatatggc aaaagcacac tcaaatagtg ttgctagaac aactttagaa
     1021 actgctgaag aacatgcaaa tagtgttgcc agaacaactt tagaaactgc tgaagaacat
     1081 gcaaataaaa aatcagctga ggcgttagca agcgctaatg tgtatgcaga cagcaagtct
     1141 tctcacacac taaaaactgc aaatagctat accgatgtga ctgtaagtaa ttcgactaag
     1201 aaagcaatcc gtgaatcgaa tcaatacaca gatcataaat tccgtcaact tgacaaccgg
     1261 ttagataagc tgtacacgga tcaattcaag agtaaaatag caggtgcaat ctggcgtaac
     1321 ttggatgaca agctcaccga ggttgtatcg cttaaagatt ttggagccaa aggtgatggt
     1381 aagacaaacg accaagatgc agtaaatgca gcgatggctt caggtaagag aattgacggt
     1441 gctggtgcta cttacaaagt atcatcttta cctgatatgg agcgattcta taacacccgc
     1501 ttcgtatggg aacgtttatc aggtcaacct ctttactatg tgagtaaagg ttttatcaat
     1561 ggtgaactct ataaaatcac ggataaccct tattacaatg cttggcctca agacaaagcg
     1621 tttgtatatg agaacgtgat atatgcacct tacatgggta gcgaccgtca tggtgttagt
     1681 cgtctgcatg tatcatgggt taagtctggt gacgatggtc aaacatggtc tacaccagag
     1741 tggttaactg atctgcatcc agattaccct acagtgaact atcattgtat gagcatgggt
     1801 gtatgtcgca accgtctgtt tgccatgatt gaaacacgta ctttagccaa gaacgcacta
     1861 accaattgtg cattgtggga tcgccctatg tctcgtagtc tgcatcttac gggtggtatc
     1921 actaaggctg caaatcagcg atatgcaaca atacatgtac cagatcacgg actattcgtg
     1981 ggcgattttg ttaacttctc taattctgcg gtaacaggtg tatcaggtga tatgactgtt
     2041 gcaacggtaa tagataagga caacttcacg gttcttacac ctaaccagca gacttcagat
     2101 ttgaataacg ctggaaagaa ttggcacatg ggtacttctt tccataagtc tccgtggcgt
     2161 aagacagatc ttggtctaat ccctcatgtc acagaggtgc atagctttgc tactattgat
     2221 aacaatggct ttgttatggg ctatcatcaa ggtgatgtag ctccacgaga agttgggctt
     2281 ttctacttcc ctgatgcttt caatagccca tctaattatg ttcgtcgtca gataccatct
     2341 gagtatgaac cagatgcggc agagccatgc atcaagtact atgacggtgt attatacctt
     2401 atcactcgtg gtactcgtgg cgaccgacta ggaagctctc tgcatcgtag tagagatata
     2461 ggtcagactt gggagtcact aagatttcca cataatgtgt atcatactac tttaccgttt
     2521 gctaaggtag gagatgacct tattatgttt ggttcagaac gtgcagaaaa tgaatgggaa
     2581 gcaggtgcac cagatgatcg ttacaaggca tcttatcctc gtaccttcta tgcacgattg
     2641 aatgtaaaca attggaatgc agctgatatt gaatgggtta acatcacaga ccaaatctat
     2701 cagggtgaca ttgtggactc tagtgtaggt gtaggttcgg tagtagttaa agacagctac
     2761 atttactata tctttggtgg cgaaaaccat ttcaacccaa tgacttatgg tgacaacaga
     2821 gataaagacc catttaaagg tcatggacac cctactgata tatactgcta taagatgcag
     2881 attgcaaatg acagtcgtgt atctcgtaag tttacatatg gtgcaactcc cggacaggct
     2941 atacctacct tcatgggtac tgatggtatt cgtactattc ctgctccact tcacttatca
     3001 ggagaggtag ttgccgcaga tatgactgtc gaacatctta ccctgaaggc aagtacaagt
     3061 gctaatattc gctcagagat gctgatggaa cgtgaatatg ggttcattgg taagactata
     3121 ccaaaggata acccaactgc ccaacgtgtc atcatctctg gtggaggggg tacagcagca
     3181 gataccggtg cgcaaattac cttacatgga gcaggttcta gtacttctag acgcgcagta
     3241 tataatgcca atgagcactt gttccaaagc ggggctatca tgccctacaa tgataacgtg
     3301 tatcctgctg gcggccctcg taatagattc actactatcc atcttacaag tgaccctatt
     3361 attacgtctg atgccactca taagtatgga tcacaaggta ttgatgagag tgtactaaag
     3421 gcatggggta aggttagctt caagcagtac aagttgattg gtgagatgtc tcgtggtgtg
     3481 caccacactc actttggtgt attggcacaa gacattgtag ctgcttttgc atccgaaggg
     3541 ttagatgcaa tcgacttcgg cattgtctcc ttcgaagaag gtcagtttgg tgttagatat
     3601 agtgaaatct taatcttgga ggctgcttat actcgttatc gtttagataa gttagaggag
     3661 atgtatgcca ctaataaaat cagttaagca agctgctgta ctccagaata cagaagaact
     3721 tattcaatca ggacgtgacc ctaagcaggc atacgccatt gccaaggatg tacaagtaaa
     3781 gcggccgatc cgcatgcgag ctcggtaccc cgggtcgacc tgcagccaag cttaattagc
     3841 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     3901 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct agcttggcga
     3961 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     4021 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     4081 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     4141 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     4201 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     4261 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     4321 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     4381 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     4441 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     4501 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     4561 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     4621 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     4681 ctggggtaat gactctctag cttgaggcat caaataaaac gaaaggctca gtcgaaagac
     4741 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4801 ccctctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4861 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4921 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     4981 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5041 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     5101 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5161 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5221 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5461 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5701 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7021 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7081 cacgaggccc tttcgtcttc ac
//
