LOCUS       Exported                6041 bp ds-DNA     circular SYN 30-NOV-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pClp-Wasabi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6041)
  AUTHORS   Dan Piraner
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 30, 2016 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6041
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        47..291
                     /note="clpP/clpX Promoter"
                     /note="From GenBank CP009072
                     
                     Promoter before ClpP/ClpX. Description found in: doi: 
                     10.1128/JB.182.23.6630-6637.2000 J. Bacteriol. December 
                     2000 vol. 182 no. 23 6630-6637"
     CDS             292..294
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             304..321
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             334..1044
                     /codon_start=1
                     /product="green variant of teal fluorescent protein mTFP1 
                     (Ai et al., 2008)"
                     /note="mWasabi"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFSYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK
                     GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA
                     VARNSTDGMDELYKG"
     terminator      1146..1193
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1230..1685
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1800..2660)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2661..2753)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      2834..3422
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3852..4043)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4615..5697)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5698..5775)
                     /gene="lacI"
                     /note="lacI promoter"
ORIGIN
        1 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaatttta cgtagcataa
       61 cgcgctaaat tcgcacaaag gcccgtcacc gccaggtggt gggctttttt ttgtcatgaa
      121 ttttgcatgg aaccgtgcga aaagcctctt tcggtgttag cgtaacaaca aaagattgtt
      181 atgcttgaaa tatggtgatg ccgtacccat aacacaggga ctagctgata atccgtccat
      241 aaggttacaa tcggtacagc aggttttttc aattttatcc aggagacgga aatgggcagc
      301 agccatcacc atcatcacca cagccaggat ccaatggtga gcaagggcga ggagaccaca
      361 atgggcgtaa tcaagcccga catgaagatc aagctgaaga tggagggcaa cgtgaatggc
      421 cacgccttcg tgatcgaggg cgagggcgag ggcaagccct acgacggcac caacaccatc
      481 aacctggagg tgaaggaggg agcccccctg cccttctcct acgacattct gaccaccgcg
      541 ttcagttacg gcaacagggc cttcaccaag taccccgacg acatccccaa ctacttcaag
      601 cagtccttcc ccgagggcta ctcttgggag cgcaccatga ccttcgagga caagggcatc
      661 gtgaaggtga agtccgacat ctccatggag gaggactcct tcatctacga gatacacctc
      721 aagggcgaga acttcccccc caacggcccc gtgatgcaga aggagaccac cggctgggac
      781 gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga agggcgacgt caagatgaag
      841 ctgctgctgg agggcggcgg ccaccaccgc gttgacttca agaccatcta cagggccaag
      901 aaggcggtga agctgcccga ctatcacttt gtggaccacc gcatcgagat cctgaaccac
      961 gacaaggact acaacaaggt gaccgtttac gagatcgccg tggcccgcaa ctccaccgac
     1021 ggcatggacg agctgtacaa gggctgagta aagaaaccgc tgctgcgaaa tttgaacgcc
     1081 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     1141 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1201 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1261 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1321 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1381 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1441 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1501 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1561 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1621 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     1681 aatttctggc ggcacgatgg catgagatta tcaaaaagga tcttcaccta gatcctttta
     1741 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1801 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1861 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1921 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1981 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2041 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2101 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2161 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2221 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2281 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2341 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2401 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2461 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2521 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2581 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2641 cggaaatgtt gaatactcat actcttcctt tttcaatcat gattgaagca tttatcaggg
     2701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggtca
     2761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2941 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3541 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3601 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3661 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3721 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3781 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3841 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3901 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     3961 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4021 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4081 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4141 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4201 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4261 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4321 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4381 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4441 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4501 ctagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     4561 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4621 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4681 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4741 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4801 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4861 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     4921 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4981 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     5041 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     5101 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     5161 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     5221 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     5281 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     5341 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5401 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5461 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5521 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5581 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5641 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5701 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     5761 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     5821 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     5881 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5941 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6001 cgccagcaac cgcacctgtg gcgccggtga tgccggccac g
//
