LOCUS       Exported                7943 bp ds-DNA     circular SYN 10-JUL-2019
DEFINITION  replaces the domain of unknown function corresponding to amino acids
            E541 -Q617 of the native EatA protein with 8H followed by an 
            in-frame XhoI induced scar of L549-E550 in the DUF deletion 
            recombinant.
ACCESSION   .
VERSION     .
KEYWORDS    pTW5126
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  AUTHORS   jmcf
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 10, 2019 from SnapGene 4.3.10
            https://www.snapgene.com
COMMENT     Alias: pBAD-EatA_N5_His_C5
FEATURES             Location/Qualifiers
     source          1..7943
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             319..4215
                     /codon_start=1
                     /label=eatA-DUF::His
                     /translation="MDKVFSLKYSFLAKGFIAVSELARRVSVKGKLKSASSIIISPITI
                     AIVSYAPPSLAATVNADISYQTFRDFAENKGAFIVGASNINIYDKNGVLVGVLDKAPMP
                     DFSSATMNTGTLPPGDHTLYSPQYVVTAKHVNGSDIMSFGHIQNNYTVVGENNHNSLDI
                     KIRRLNKIVTEVAPAEISSVGAVNGAYQEGGRFKAFYRLGGGLQYIKDKNGNLTPVYTN
                     GGFLTGGTISALSSYNNGQMITAPTGDIFNPANGPLANYLNKGDSGSPLFAYDSLDKKW
                     VLVGVLSSGSEHGNNWVVTTQDFLHQQPKHDFDKTISYDSEKGSLQWRYNKNSGVGTLS
                     QESVVWDMHGKKGGDLNAGKNLQFTGNNGEIILHDSIDQGAGYLQFFDNYTVTSLTDQT
                     WTGGGIITEKGVNVLWQVNGVNDDNLHKVGEGTLTVNGKGVNNGGLKVGDGTVILNQRP
                     DDNGHKQAFSSINISSGRATVILSDANQVNPDKISWGYRGGTLDLNGNNVNFTRLQAAD
                     YGAIVSNNNKNKSELTLKLQTLNHHHHHHHHLEQPYMFHGQLNGYMDVNIHPLSGKDVL
                     TLDGSVNLPEGVITKKSGTLIFQGHPVIHAGMTTSAGQSDWENRQFTMDKLRLDAATFH
                     LSRNAHMQGDISAANGSTVILGSSRVFTDKNDGTGNAVSSVEGSSIATTAGDQSYYSGN
                     VLLENHSSLEVRENFTGGIEAYDSSVSVTSQNAIFDHVGSFVNSSLLLEKGAKLTAQSG
                     IFTNNTMKIKENASLTLTGIPSVGKPGYYSPVTSTTEGIHLGERASLSVKNMGYLSSNI
                     TAENSAAIINLGDSNATIGKTDSPLFSTLMRGYNAVLQGNIMGPQSSVNMNNALWHSDR
                     NSELKELKANDSQIELGVRGHFAKLRVKELIASNSVFLVHANNSQADQLNVTDKLQGSN
                     NTILVDFFNKAANGTNVTLITAPKGSDENTFKAGTQQIGFSNITPEIRTENTDTATQWV
                     LTGYQSVADARASKIATDFMDSGYKSFLTEVNNLNKRMGDLRDSQGDAGGWARIMNGTG
                     SGESGYRDNYTHVQIGADRKHELNGIDLFTGALLTYTDNNASSQAFSGKTKSLGGGVYA
                     SGLFESGAYFDLIGKYLHHDNRYTLNFASLGERSYTSHSLYAGAEIGYRYHMSENTWVE
                     PQMELVYGSVSGKSFNWKDQGMQLSMKDKDYHPLIGRTGVDVGRAFSGDTWKVTVRAGL
                     GYQFDLLANGETVLQDASGKKHFKGEKDSRMLMNVGTNVEVKDNMRFGLELEKSAFGRY
                     NIDNSINANFRYYF"
     primer_bind     319..347
                     /label=jf110414.1 5-Nterm
     primer_bind     complement(1922..1968)
                     /label=jf061615.3 DUFhis3'
     CDS             1939..1962
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     1963..1990
                     /label=jf061615.2 DUFdel5'
     primer_bind     complement(4193..4221)
                     /label=jf022515.7 C3-3
     CDS             4226..4255
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4271..4288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4514..4600
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4692..4719
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4738..4829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4830..5690
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5861..6449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6635..6775
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(7039..7917)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggataaagtg ttctctctta agtatagttt tttagccaaa
      361 ggttttattg ctgtttctga acttgcccgt cgcgtttctg ttaaagggaa actgaagagt
      421 gcttcatcaa taattatttc accaataaca attgctattg tttcttatgc acccccatct
      481 cttgctgcaa cagttaatgc agatatatcg tatcaaacat ttcgggattt tgccgaaaat
      541 aaaggagctt ttatagttgg cgcatcaaat ataaatatct acgataagaa tggagtgtta
      601 gttggagtgc ttgataaagc tccaatgcct gattttagta gcgccacgat gaatacaggg
      661 acattaccac caggagacca tacactgtac tcacctcaat atgttgtcac agcaaagcat
      721 gttaatggat cagatataat gagttttgga catattcaaa ataattatac tgtagtagga
      781 gagaacaacc ataatagcct tgatattaaa atacggcgtt taaataagat tgtcacggag
      841 gtcgcccctg cagaaatctc cagtgttgga gctgtaaatg gcgcttatca agaagggggg
      901 cgttttaaag ccttttatag gcttggaggt ggattgcaat atataaagga taaaaatggg
      961 aatcttacac cggtatatac aaatggtggt ttcctaaccg gaggtactat cagtgcttta
     1021 agctcatata acaatggcca aatgattacc gcacctacag gcgatatatt taatccagct
     1081 aatgggcctc ttgcaaacta tctaaataaa ggtgatagtg gctctccttt atttgcgtat
     1141 gactctctgg acaaaaaatg ggttttagtt ggcgttctgt catcaggaag cgagcatggt
     1201 aataactggg ttgtcacaac tcaagatttt cttcatcagc aaccaaaaca tgattttgat
     1261 aaaacaatat catatgactc tgaaaagggt agcttacaat ggagatataa taaaaattca
     1321 ggagtgggaa cattaagtca agagagcgtt gtgtgggaca tgcatggaaa aaaaggggga
     1381 gatctaaacg caggtaaaaa tcttcaattt acaggaaata atggagagat tattttacac
     1441 gactctatag atcaaggggc tggctatttg cagttttttg acaactacac agttacatcc
     1501 ttaactgacc aaacatggac cggaggtggt atcattactg aaaaaggtgt aaatgtgctt
     1561 tggcaggtta atggtgttaa tgatgataac ttacataaag ttggtgaagg cacattaact
     1621 gttaatggaa aaggggttaa taatggagga ctgaaagtcg gtgatggaac cgtaattctg
     1681 aatcaacgcc ctgatgataa tggacacaag caagccttta gctctattaa catttccagt
     1741 ggtcgtgcaa cagttatact ttcagatgct aatcaagtta acccagataa aatatcatgg
     1801 ggatatagag gcggtactct tgatttaaat ggaaataatg taaactttac tcgtcttcag
     1861 gctgcagatt atggtgctat tgtttctaac aataacaaaa acaaatctga attaacactt
     1921 aaattacaaa cactaaatca tcatcatcat catcatcatc atctcgagca accttatatg
     1981 tttcatggac aacttaatgg ttatatggat gtaaatatac atccactctc tggtaaggat
     2041 gtgctgactc ttgatggttc tgttaatctg cctgaagggg tgataacgaa aaagtcaggt
     2101 actctgatat ttcaagggca tccggtgatt catgctggaa tgacaacctc agccggccag
     2161 agtgattggg aaaatcgtca gtttacaatg gataaactga ggcttgatgc agcaacattc
     2221 catctctcca gaaatgctca tatgcaggga gatattagtg ctgccaacgg aagcaccgtc
     2281 attctgggaa gttctcgggt ctttactgac aagaatgacg gaaccggtaa tgcggtatct
     2341 tctgttgaag ggagttccat tgcaacaaca gctggtgacc aaagttatta cagcggtaat
     2401 gtgctgctgg aaaaccattc gtctctagag gtcagggaga attttactgg tggtattgag
     2461 gcttatgaca gttctgttag tgtgacctct cagaatgcta tttttgacca tgttggtagc
     2521 tttgttaata gtagtctgct tctcgaaaaa ggagcaaaac tgacagcaca gagtggtatt
     2581 ttcacaaata acactatgaa aataaaagaa aacgcctccc tgactctgac agggatacct
     2641 tctgtaggaa agccagggta ttattcacct gtgacctcga ctactgaagg aattcatctc
     2701 ggtgagcgag ccagcctttc agtgaaaaat atgggctatc tgagttcaaa tattacagca
     2761 gagaactctg cagcaattat taatctggga gacagtaatg caactatcgg gaagacggac
     2821 tctccattat tcagtacctt aatgagggga tataatgctg ttttgcaggg caatattatg
     2881 gggccccaga gctcagtgaa tatgaacaat gctctgtggc actctgatag aaattcggaa
     2941 ctcaaagagc tgaaagccaa cgactcccaa atagagttgg gtgtaagagg gcattttgca
     3001 aaactgcggg taaaagagct tattgcgtct aactcagtgt ttcttgtaca tgcaaacaat
     3061 agccaggctg accagttgaa cgttaccgac aaactgcagg gcagcaacaa tactattctt
     3121 gttgactttt ttaacaaagc agccaatggt acaaatgtga cgttaattac tgcaccaaaa
     3181 ggcagtgatg aaaatacatt caaagccgga acccagcaga ttggattcag taatatcacg
     3241 ccagaaatca ggacagaaaa tacggataca gccacacagt gggtgctgac tggatatcag
     3301 tctgtcgctg atgccagagc ctcgaaaatc gcaacggact ttatggattc aggttataaa
     3361 tctttcctga cggaagtcaa taatctgaac aaacgtatgg gagatttacg ggatagtcag
     3421 ggagatgctg gagggtgggc gcgtatcatg aatggtaccg gttcaggtga gagtggttac
     3481 agagataact atacccacgt tcagattggt gcagacagaa agcatgagct gaacggtata
     3541 gatttattca ccggtgcatt actgacttat acagacaaca atgctagcag ccaggctttc
     3601 agcggtaaaa caaaatcgct agggggaggg gtgtatgcat caggtctctt tgagtctgga
     3661 gcttattttg acctgattgg taaatatctc catcatgata atcggtatac gttgaatttt
     3721 gcctccttgg gggaaagaag ctacacctcc cattctttgt atgctggagc tgaaatcggg
     3781 tatcgttatc acatgtcaga aaatacatgg gtggaaccac agatggaact ggtttatggt
     3841 tcggtatcag gaaagtcatt taactggaaa gaccagggaa tgcaactgag tatgaaagac
     3901 aaagactatc acccactaat tggtcgaaca ggtgtggatg taggtagagc gttctctgga
     3961 gatacctgga aagtaacagt acgtgcagga ctgggttacc agttcgattt gctggcaaat
     4021 ggagaaactg ttttacagga tgcttctggt aaaaaacact tcaaaggtga aaaagacagc
     4081 aggatgctaa tgaacgtggg gacgaatgtg gaagttaaag acaatatgcg ttttggtctg
     4141 gagttggaga agtcggcgtt tgggagatat aacatagaca actctataaa tgctaacttc
     4201 cgttattatt tctgaaagct ttctagaaca aaaactcatc tcagaagagg atctgaatag
     4261 cgccgtcgac catcatcatc atcatcattg agtttaaacg gtctccagct tggctgtttt
     4321 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     4381 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     4441 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     4501 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     4561 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     4621 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     4681 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     4741 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4801 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4861 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4921 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4981 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5041 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     5101 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5161 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5221 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5281 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5341 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5401 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5461 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5521 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5581 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5641 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5701 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5761 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5821 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5881 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5941 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6001 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6061 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6541 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6601 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     6661 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     6721 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     6781 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6841 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6901 gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg cataatgtgc
     6961 ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat
     7021 tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca
     7081 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
     7141 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     7201 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     7261 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     7321 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     7381 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
     7441 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     7501 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     7561 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     7621 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     7681 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
     7741 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     7801 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     7861 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     7921 tttcatactc ccgccattca gag
//
