LOCUS       Exported                5832 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pNeae2-complete
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 9, 2021 from SnapGene Viewer 5.2.1
            https://www.snapgene.com
COMMENT     LOCUS Exported 5832 bp ds-DNA circular SYN 21-ABR-2016 DEFINITION 
            synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAK-Backbone1 
            SOURCE synthetic DNA construct ORGANISM synthetic DNA construct 
            REFERENCE 1 (bases 1 to 5832) AUTHORS . TITLE Direct Submission 
            JOURNAL Exported 8 de may de 2016 from SnapGene Viewer 3.0.2 
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5832
                     /organism="synthetic DNA construct"
                     /label=synthetic DNA construct
                     /note="synthetic DNA construct"
     promoter        5..82
                     /note="lacI promoter"
     CDS             83..1165
                     /codon_start=1
                     /label=lacI repressor
                     /note="lacI repressor"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1233..1254
                     /note="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in thepresence  
                     of cAMP."
     promoter        1269..1299
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1307..1323
                     /note="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1331..1347
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1383..3464
                     /codon_start=1
                     /label=Neae2
                     /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
                     KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
                     APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
                     NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
                     FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
                     FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
                     YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
                     DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
                     PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
                     VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
                     TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
                     AKTAEMTSALNASAVIFFDGAPVPYPDPLEPAQPAMAHHHHHHAAAEQKLISEEDAAA"
     misc_feature    1383..3344
                     /label=N-terminal fragment Intimin
                     /note="N-terminal fragment Intimin"
                     /note="N-terminal fragment Intimin (Neae)"
     misc_feature    2677..2699
                     /label=Oligo eae-3
                     /note="Oligo eae-3"
     misc_feature    2982..3008
                     /label=eae4
                     /note="eae4"
     misc_feature    3345..3383
                     /label=E-tag
                     /note="E-tag"
     misc_feature    3399..3416
                     /label=6xHis
                     /note="6xHis"
     misc_feature    3426..3452
                     /label=myc
                     /note="myc"
     rep_origin      3551..4006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
     CDS             4193..4852
                     /codon_start=1
                     /label=cat
                     /note="cat"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      5184..5772
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
       61 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
      121 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
      181 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
      241 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
      301 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
      361 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
      421 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
      481 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
      541 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
      601 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
      661 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
      721 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
      781 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
      841 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata
      901 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
      961 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     1021 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     1081 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1141 ttcccgactg gaaagcgggc agtgagcggt acccgataaa agcggcttcc tgacaggagg
     1201 ccgttttgtt ttgcagccca cctcaacgca attaatgtga gttagctcac tcattaggca
     1261 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     1321 caatttcaca caggaaacag ctatgaccat gattacgaat ttctagataa cgagggcaaa
     1381 tcatgattac tcatggttgt tatacccgga cccggcacaa gcataagcta aaaaaaacat
     1441 tgattatgct tagtgctggt ttaggattgt ttttttatgt taatcagaat tcatttgcaa
     1501 atggtgaaaa ttattttaaa ttgggttcgg attcaaaact gttaactcat gatagctatc
     1561 agaatcgcct tttttatacg ttgaaaactg gtgaaactgt tgccgatctt tctaaatcgc
     1621 aagatattaa tttatcgacg atttggtcgt tgaataagca tttatacagt tctgaaagcg
     1681 aaatgatgaa ggccgcgcct ggtcagcaga tcattttgcc actcaaaaaa cttccctttg
     1741 aatacagtgc actaccactt ttaggttcgg cacctcttgt tgctgcaggt ggtgttgctg
     1801 gtcacacgaa taaactgact aaaatgtccc cggacgtgac caaaagcaac atgaccgatg
     1861 acaaggcatt aaattatgcg gcacaacagg cggcgagtct cggtagccag cttcagtcgc
     1921 gatctctgaa cggcgattac gcgaaagata ccgctcttgg tatcgctggt aaccaggctt
     1981 cgtcacagtt gcaggcctgg ttacaacatt atggaacggc agaggttaat ctgcagagtg
     2041 gtaataactt tgacggtagt tcactggact tcttattacc gttctatgat tccgaaaaaa
     2101 tgctggcatt tggtcaggtc ggagcgcgtt acattgactc ccgctttacg gcaaatttag
     2161 gtgcgggtca gcgttttttc cttcctgcaa acatgttggg ctataacgtc ttcattgatc
     2221 aggatttttc tggtgataat acccgtttag gtattggtgg cgaatactgg cgagactatt
     2281 tcaaaagtag cgttaacggc tatttccgca tgagcggctg gcatgagtca tacaataaga
     2341 aagactatga tgagcgccca gcaaatggct tcgatatccg ttttaatggc tatctaccgt
     2401 catatccggc attaggcgcc aagctgatat atgagcagta ttatggtgat aatgttgctt
     2461 tgtttaattc tgataagctg cagtcgaatc ctggtgcggc gaccgttggt gtaaactata
     2521 ctccgattcc tctggtgacg atggggatcg attaccgtca tggtacgggt aatgaaaatg
     2581 atctccttta ctcaatgcag ttccgttatc agtttgataa atcgtggtct cagcaaattg
     2641 aaccacagta tgttaacgag ttaagaacat tatcaggcag ccgttacgat ctggttcagc
     2701 gtaataacaa tattattctg gagtacaaga agcaggatat tctttctctg aatattccgc
     2761 atgatattaa tggtactgaa cacagtacgc agaagattca gttgatcgtt aagagcaaat
     2821 acggtctgga tcgtatcgtc tgggatgata gtgcattacg cagtcagggc ggtcagattc
     2881 agcatagcgg aagccaaagc gcacaagact accaggctat tttgcctgct tatgtgcaag
     2941 gtggcagcaa tatttataaa gtgacggctc gcgcctatga ccgtaatggc aatagctcta
     3001 acaatgtaca gcttactatt accgttctgt cgaatggtca agttgtcgac caggttgggg
     3061 taacggactt tacggcggat aagacttcgg ctaaagcgga taacgccgat accattactt
     3121 ataccgcgac ggtgaaaaag aatggggtag ctcaggctaa tgtccctgtt tcatttaata
     3181 ttgtttcagg aactgcaact cttggggcaa atagtgccaa aacggatgct aacggtaagg
     3241 caaccgtaac gttgaagtcg agtacgccag gacaggtcgt cgtgtctgct aaaaccgcgg
     3301 agatgacttc agcacttaat gccagtgcgg ttatattttt tgatggtgcg ccggtgccgt
     3361 atccggatcc gctggaaccg gcccagccgg ccatggcgca tcaccatcac catcacgcgg
     3421 ccgccgaaca aaaactcatc tcagaagagg atgcagctgc ataaaagctt gacctgtgaa
     3481 gtgaaaaatg gcgcacattg tgcgacattt tttttgtctg ccgtttaccg ctactgcgtc
     3541 acggatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3601 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3661 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     3721 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3781 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3841 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3901 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3961 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt
     4021 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4081 ccgctcatgt cgagacgttg ggtgaggttc caactttcac cataatgaaa taagatcact
     4141 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     4201 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     4261 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     4321 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     4381 tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt
     4441 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     4501 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     4561 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     4621 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     4681 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     4741 gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc atgtcggcag
     4801 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     4861 aggcagttat tggtgccctt aaacgcctgg tgctacgcct gaataagtga taataagcgg
     4921 atgaatggca gaaattcgaa agcaaattcg acccggtcgt cggttcaggg cagggtcgtt
     4981 aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga agcagtgtga
     5041 ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg gaggtaataa
     5101 ttgctcgaca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5161 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5221 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5281 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5341 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5401 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5461 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5521 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5581 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5641 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5701 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5761 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5821 tttgctcaca tg
//
