LOCUS       Exported                7051 bp DNA     circular SYN 05-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7051)
  AUTHORS   Finne lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     host strain M15 pREP4
FEATURES             Location/Qualifiers
     source          1..7051
                     /organism="recombinant plasmid"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        5..49
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    25..41
                     /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    57..73
                     /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             92..103
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             110..112
                     /codon_start=1
                     /label=start codon
                     /translation="M"
     CDS             122..139
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             149..820
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)
                     >EU827527.1:4-675 Synthetic construct DsRed-Monomer gene, 
                     complete cds"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="DNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKL
                     QVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVV
                     EVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHA
                     LKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSG
                     SQ"
     CDS             827..1216
                     /codon_start=1
                     /product=">gi|48606|emb|X13882.1| Y.enterocolitica yadA 
                     gene for YadA "
                     /label=yadA stalk
                     /translation="GIANNYTDSKSAETLENARKEAFAQSKDVLNMAKAHSNSVARTTL
                     ETAEEHANSVARTTLETAEEHANKKSAEALASANVYADSKSSHTLKTANSYTDVTVSNS
                     TKKAIRESNQYTDHKFRQLDNRLDKL"
     CDS             1229..3631
                     /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    2444..2446
                     /label=H417Y
                     /note="CAT -> TAT (H417->Y) "
     misc_feature    2660..2662
                     /label=N489D
                     /note="AAC -> GAC (N489->D) "
     terminator      3793..3887
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3931..4590
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      4655..4741
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4896..5036
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5222..5810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5981..6841)
                     /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(6842..6946)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaatcataa aaaatttatt tgctttgtga gcggataaca attataatag attcaattgt
       61 gagcggataa caatttcaca cagaattcat taaagaggag aaattaacta tgagaggatc
      121 tcaccatcac catcaccatg gcggccgcga caacaccgag gacgtcatca aggagttcat
      181 gcagttcaag gtgcgcatgg agggctccgt gaacggccac tacttcgaga tcgagggcga
      241 gggcgagggc aagccctacg agggcaccca gaccgccaag ctgcaggtga ccaagggcgg
      301 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggccta
      361 cgtgaagcac cccgccgaca tccccgacta catgaagctg tccttccccg agggcttcac
      421 ctgggagcgc tccatgaact tcgaggacgg cggcgtggtg gaggtgcagc aggactcctc
      481 cctgcaggac ggcaccttca tctacaaggt gaagttcaag ggcgtgaact tccccgccga
      541 cggccccgta atgcagaaga agactgccgg ctgggagccc tccaccgaga agctgtaccc
      601 ccaggacggc gtgctgaagg gcgagatctc ccacgccctg aagctgaagg acggcggcca
      661 ctacacctgc gacttcaaga ccgtgtacaa ggccaagaag cccgtgcagc tgcccggcaa
      721 ccactacgtg gactccaagc tggacatcac caaccacaac gaggactaca ccgtggtgga
      781 gcagtacgag cacgccgagg cccgccactc cggctcccag actagtggga tcgcaaataa
      841 ctatactgat agtaaaagtg ctgaaacatt ggaaaatgcg cgtaaagagg cttttgctca
      901 gtctaaagat gttttgaata tggcaaaagc acactcaaat agtgttgcta gaacaacttt
      961 agaaactgct gaagaacatg caaatagtgt tgccagaaca actttagaaa ctgctgaaga
     1021 acatgcaaat aaaaaatcag ctgaggcgtt agcaagcgct aatgtgtatg cagacagcaa
     1081 gtcttctcac acactaaaaa ctgcaaatag ctataccgat gtgactgtaa gtaattcgac
     1141 taagaaagca atccgtgaat cgaatcaata cacagatcat aaattccgtc aacttgacaa
     1201 ccggttagat aagctgtaca cggatcaatt caagagtaaa atagcaggtg caatctggcg
     1261 taacttggat gacaagctca ccgaggttgt atcgcttaaa gattttggag ccaaaggtga
     1321 tggtaagaca aacgaccaag atgcagtaaa tgcagcgatg gcttcaggta agagaattga
     1381 cggtgctggt gctacttaca aagtatcatc tttacctgat atggagcgat tctataacac
     1441 ccgcttcgta tgggaacgtt tatcaggtca acctctttac tatgtgagta aaggttttat
     1501 caatggtgaa ctctataaaa tcacggataa cccttattac aatgcttggc ctcaagacaa
     1561 agcgtttgta tatgagaacg tgatatatgc accttacatg ggtagcgacc gtcatggtgt
     1621 tagtcgtctg catgtatcat gggttaagtc tggtgacgat ggtcaaacat ggtctacacc
     1681 agagtggtta actgatctgc atccagatta ccctacagtg aactatcatt gtatgagcat
     1741 gggtgtatgt cgcaaccgtc tgtttgccat gattgaaaca cgtactttag ccaagaacgc
     1801 actaaccaat tgtgcattgt gggatcgccc tatgtctcgt agtctgcatc ttacgggtgg
     1861 tatcactaag gctgcaaatc agcgatatgc aacaatacat gtaccagatc acggactatt
     1921 cgtgggcgat tttgttaact tctctaattc tgcggtaaca ggtgtatcag gtgatatgac
     1981 tgttgcaacg gtaatagata aggacaactt cacggttctt acacctaacc agcagacttc
     2041 agatttgaat aacgctggaa agaattggca catgggtact tctttccata agtctccgtg
     2101 gcgtaagaca gatcttggtc taatccctca tgtcacagag gtgcatagct ttgctactat
     2161 tgataacaat ggctttgtta tgggctatca tcaaggtgat gtagctccac gagaagttgg
     2221 gcttttctac ttccctgatg ctttcaatag cccatctaat tatgttcgtc gtcagatacc
     2281 atctgagtat gaaccagatg cggcagagcc atgcatcaag tactatgacg gtgtattata
     2341 ccttatcact cgtggtactc gtggcgaccg actaggaagc tctctgcatc gtagtagaga
     2401 tataggtcag acttgggagt cactaagatt tccacataat gtgtatcata ctactttacc
     2461 gtttgctaag gtaggagatg accttattat gtttggttca gaacgtgcag aaaatgaatg
     2521 ggaagcaggt gcaccagatg atcgttacaa ggcatcttat cctcgtacct tctatgcacg
     2581 attgaatgta aacaattgga atgcagctga tattgaatgg gttaacatca cagaccaaat
     2641 ctatcagggt gacattgtgg actctagtgt aggtgtaggt tcggtagtag ttaaagacag
     2701 ctacatttac tatatctttg gtggcgaaaa ccatttcaac ccaatgactt atggtgacaa
     2761 cagagataaa gacccattta aaggtcatgg acaccctact gatatatact gctataagat
     2821 gcagattgca aatgacagtc gtgtatctcg taagtttaca tatggtgcaa ctcccggaca
     2881 ggctatacct accttcatgg gtactgatgg tattcgtact attcctgctc cacttcactt
     2941 atcaggagag gtagttgccg cagatatgac tgtcgaacat cttaccctga aggcaagtac
     3001 aagtgctaat attcgctcag agatgctgat ggaacgtgaa tatgggttca ttggtaagac
     3061 tataccaaag gataacccaa ctgcccaacg tgtcatcatc tctggtggag ggggtacagc
     3121 agcagatacc ggtgcgcaaa ttaccttaca tggagcaggt tctagtactt ctagacgcgc
     3181 agtatataat gccaatgagc acttgttcca aagcggggct atcatgccct acaatgataa
     3241 cgtgtatcct gctggcggcc ctcgtaatag attcactact atccatctta caagtgaccc
     3301 tattattacg tctgatgcca ctcataagta tggatcacaa ggtattgatg agagtgtact
     3361 aaaggcatgg ggtaaggtta gcttcaagca gtacaagttg attggtgaga tgtctcgtgg
     3421 tgtgcaccac actcactttg gtgtattggc acaagacatt gtagctgctt ttgcatccga
     3481 agggttagat gcaatcgact tcggcattgt ctccttcgaa gaaggtcagt ttggtgttag
     3541 atatagtgaa atcttaatct tggaggctgc ttatactcgt tatcgtttag ataagttaga
     3601 ggagatgtat gccactaata aaatcagtta agcaagctgc tgtactccag aatacagaag
     3661 aacttattca atcaggacgt gaccctaagc aggcatacgc cattgccaag gatgtacaag
     3721 taaagcggcc gatccgcatg cgagctcggt accccgggtc gacctgcagc caagcttaat
     3781 tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     3841 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagcttg
     3901 gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     3961 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     4021 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     4081 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     4141 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     4201 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     4261 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     4321 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     4381 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     4441 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     4501 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     4561 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
     4621 acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa
     4681 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     4741 tccgccctct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4801 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4861 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     4921 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4981 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     5041 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5101 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5161 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5641 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6001 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6061 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6121 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6181 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6241 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6301 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6361 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6421 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6481 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6541 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6601 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6661 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6721 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6781 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6841 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6901 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6961 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7021 gtatcacgag gccctttcgt cttcacctcg a
//
