LOCUS       62056_3725        7386 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             458..1249
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1657..1678
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1693..1723
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /label=lac operator
                     /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)."
     primer_bind     1755..1771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1792..1810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(1829..1856)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(1881..4541)
                     /codon_start=1
                     /label=NgAgo
                     /note="Argonaute protein from Natronobacterium gregoryi
                     (Gao et al., 2016)"
                     /translation="MTVIDLDSTTTADELTSGHTYDISVTLTGVYDNTDEQHPRMSLAF
                     EQDNGERRYITLWKNTTPKDVFTYDYATGSTYIFTNIDYEVKDGYENLTATYQTTVENA
                     TAQEVGTTDEDETFAGGEPLDHHLDDALNETPDDAETESDSGHVMTSFASRDQLPEWTL
                     HTYTLTATDGAKTDTEYARRTLAYTVRQELYTDHDAAPVATDGLMLLTPEPLGETPLDL
                     DCGVRVEADETRTLDYTTAKDRLLARELVEEGLKRSLWDDYLVRGIDEVLSKEPVLTCD
                     EFDLHERYDLSVEVGHSGRAYLHINFRHRFVPKLTLADIDDDNIYPGLRVKTTYRPRRG
                     HIVWGLRDECATDSLNTLGNQSVVAYHRNNQTPINTDLLDAIEAADRRVVETRRQGHGD
                     DAVSFPQELLAVEPNTHQIKQFASDGFHQQARSKTRLSASRCSEKAQAFAERLDPVRLN
                     GSTVEFSSEFFTGNNEQQLRLLYENGESVLTFRDGARGAHPDETFSKGIVNPPESFEVA
                     VVLPEQQADTCKAQWDTMADLLNQAGAPPTRSETVQYDAFSSPESISLNVAGAIDPSEV
                     DAAFVVLPPDQEGFADLASPTETYDELKKALANMGIYSQMAYFDRFRDAKIFYTRNVAL
                     GLLAAAGGVAFTTEHAMPGDADMFIGIDVSRSYPEDGASGQINIAATATAVYKDGTILG
                     HSSTRPQLGEKLQSTDVRDIMKNAILGYQQVTGESPTHIVIHRDGFMNEDLDPATEFLN
                     EQGVEYDIVEIRKQPQTRLLAVSDVQYDTPVKSIAAINQNEPRATVATFGAPEYLATRD
                     GGGLPRPIQIERVAGETDIETLTRQVYLLSQSHIQVHNSTARLPITTAYADQASTHATK
                     GYLVQTGAFESNVGFL"
     RBS             complement(4558..4580)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(4595..4623)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(4935..5594)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(5595..6364)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
     1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1801 tcactaaagg gaacaaaagc tgggtaccaa aaggccatcc gtcaggatgg ccttctgtcg
     1861 acggtatcga tctcgagtta gaggaatccg acattagact cgaacgctcc ggtctggacg
     1921 aggtaaccct tggtcgcgtg agtacttgcc tggtcggcgt atgcggtggt gatgggtagg
     1981 cgcgcagtcg agttatggac ctggatatgc gactgggaga gcagatagac ttggcgagtg
     2041 agcgtctcga tgtcggtttc gccggcgact cgttcaattt ggattgggcg gggaaggccg
     2101 cctccatcgc gtgtcgctaa gtattcgggt gcgccgaagg tggcgaccgt tgcccgtggc
     2161 tcgttctggt tgatagcggc gatgctcttc acaggcgtat cgtactgcac atcggagact
     2221 gccagcaggc gtgtctgggg ctgcttgcgg atttcgacga tgtcgtactc gacgccttgt
     2281 tcgttgagga attccgtggc ggggtcgagg tcttcgttca tgaagccatc acggtggatg
     2341 acgatatggg tcggcgactc accggtcacc tgctggtagc cgaggatggc attcttcata
     2401 atgtcacgaa catccgtcga ctgtagtttc tccccgagct gcggtcgggt ggacgagtgg
     2461 ccgaggatag ttccatcctt gtagacggcg gtcgccgtcg cggcaatgtt tatctggccg
     2521 ctggcaccgt cctcggggta gctccgagag acatcaatcc caatgaacat atctgcgtcc
     2581 ccaggcatcg catgttcggt tgtgaatgcg acgccgccag cggctgccag cagcccgagt
     2641 gccacgttac gagtatagaa tattttcgcg tcgcggaacc ggtcgaagta cgccatctgg
     2701 ctgtaaatgc ccatgttggc aagcgccttc ttcagctcgt cgtacgtctc tgtcggactg
     2761 gcgaggtctg cgaatccttc ttggtccggc ggcagtacga cgaatgccgc gtctacctcg
     2821 ctagggtcga tggctccagc cacattgagg ctgatgctct ctggcgagga gaacgcatca
     2881 tattggacgg tctcgctccg tgtcggtggc gcgccagctt ggttgaggag gtcagccatc
     2941 gtgtcccact gcgctttgca ggtatctgcc tgctgctcgg gcagtactac ggccacctcg
     3001 aacgactctg gtggattgac gatacctttc gagaatgtct cgtcggggtg cgcaccacgc
     3061 gccccgtcgc ggaacgtcag aaccgactca ccgttctcgt agaggaggcg cagttgctgc
     3121 tcgttgttcc cggtgaaaaa ctccgaggag aactctaccg tggacccatt gagacgcacc
     3181 gggtcaagcc gctcggcgaa cgcttgcgct ttctcgctgc agcgggaggc cgagagacgc
     3241 gtctttgagc gggcctgttg gtggaatccg tcggaggcga actgcttaat ttggtgcgta
     3301 ttcggttcga ccgcaagcag ttcttggggg aatgagacag catcatcgcc gtgcccttga
     3361 cgtcgggttt cgacgactcg ccggtcagcg gcctcgatag cgtcgaggag gtcagtgtta
     3421 ataggtgtct gattgttgcg gtggtatgca acgacggact ggtttcccag cgtgttgagc
     3481 gagtcggtgg cgcactcgtc ccgcagaccc cagacgatat gtcctcgccg ggggcgatac
     3541 gtcgtcttca cccggagccc aggatagatg ttgtcatcat cgatgtctgc gagcgtcagc
     3601 ttcggtacga accggtggcg gaagttgatg tgaaggtacg cccgcccact gtgaccgact
     3661 tcgacagaga ggtcataccg ctcatgtagg tcgaactcat cgcaagtcag cacaggctcc
     3721 tttgagagga cttcatcgat gccgcgaacg aggtagtcat cccagaggga gcgtttgagc
     3781 ccctcttcga cgagttcgcg ggcgagtaac cggtctttgg ccgtggtgta atcgagtgtc
     3841 cgagtctcgt ccgcctcgac ccggacaccg caatcgaggt caagcggggt ctcgccgagc
     3901 ggctctggcg tgagaagcat tagcccgtca gttgcaaccg gagccgcatc atggtcggta
     3961 tagagttcct gccgtaccgt gtatgcgagg gttcgtcgcg cgtactccgt gtccgtcttt
     4021 gcgccgtctg tggctgttag cgtatacgta tgcagcgtcc actcagggag ttggtcgcgg
     4081 gaggcgaacg aggtcatcac atggcctgag tcgctctctg tctccgcgtc gtctggcgtc
     4141 tcattgagcg cgtcgtccaa gtgatggtcg agcggctcgc cgcccgcgaa cgtttcgtcc
     4201 tcgtcagtcg tcccgacttc ctgagcggta gcgttctcga cggtcgtctg gtatgttgca
     4261 gtcagattct cgtagccgtc cttcacttcg tagtcgatgt tagtgaagat gtacgtcgag
     4321 cccgtggcgt agtcgtatgt aaagacatcc ttgggtgtcg tgttcttcca cagggtaatg
     4381 taacgccgct cgccgttgtc ctgctcgaat gcgagagaca tgcgaggatg ctgctcgtcg
     4441 gtgttatcgt agacaccggt gagcgtgact gagatgtcgt acgtgtgtcc cgatgtcagt
     4501 tcgtctgcgg tggtggtcga atcgaggtca atcactgtca taagcttcca tggtatatct
     4561 ccttcttaaa gttaaacaaa attatttcta gaggcattat acgagccgat gattaattgt
     4621 caaggatcca ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc
     4681 tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat
     4741 tctgcctccc agagcctgat aaaaacggtg aatccgttag cgaggtgccg ccggcttcca
     4801 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     4861 atagggcggc gaggcggcta cagccgatag tctggaacag cgcacttacg ggttgctgcg
     4921 caacccaagt gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa cggctcgcca
     4981 tcgtccagaa aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat
     5041 tcctccgttt cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg
     5101 ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg
     5161 cggcgcaggt cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg
     5221 gcggcactga acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca
     5281 ttgaccacgt aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc
     5341 tcggccacca agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg
     5401 gaaagtgtct tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga
     5461 tgcagcagca ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct
     5521 ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg
     5581 gactgcgtgg ccatgcttat ctccatgcgg tagggtgccg cacggttgcg gcaccatgcg
     5641 caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt
     5701 caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc gcaatgagct
     5761 gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt cgccctatat
     5821 ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc cttcggcgcg
     5881 gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc gcggccttcg
     5941 gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg tgaggctcgg
     6001 ggggcaggcg ggcgggcttc gccttcgact gcccccactc gcataggctt gggtcgttcc
     6061 aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc gccttccact
     6121 tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa
     6181 aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg tgggtatgtg
     6241 gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc
     6301 tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc cagccggtgg
     6361 ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc gaccgcgcag
     6421 ggcgaagccc ggagagcaag cccgtagggc gccgcagccg ccgtaggcgg tcacgacttt
     6481 gcgaagcaaa gtctagtgag tatactcaag cattgagtgg cccgccggag gcaccgcctt
     6541 gcgctgcccc cgtcgagccg gttggacacc aaaagggagg ggcaggcatg gcggcatacg
     6601 cgatcatgcg atgcaagaag ctggcgaaaa tgggcaacgt ggcggccagt ctcaagcacg
     6661 cctaccgcga gcgcgagacg cccaacgctg acgccagcag gacgccagag aacgagcact
     6721 gggcggccag cagcaccgat gaagcgatgg gccgactgcg cgagttgctg ccagagaagc
     6781 ggcgcaagga cgctgtgttg gcggtcgagt acgtcatgac ggccagcccg gaatggtgga
     6841 agtcggccag ccaagaacag caggcggcgt tcttcgagaa ggcgcacaag tggctggcgg
     6901 acaagtacgg ggcggatcgc atcgtgacgg ccagcatcca ccgtgacgaa accagcccgc
     6961 acatgaccgc gttcgtggtg ccgctgacgc aggacggcag gctgtcggcc aaggagttca
     7021 tcggcaacaa agcgcagatg acccgcgacc agaccacgtt tgcggccgct gtggccgatc
     7081 tagggctgca acggggcatc gagggcagca aggcacgtca cacgcgcatt caggcgttct
     7141 acgaggccct ggagcggcca ccagtgggcc acgtcaccat cagcccgcaa gcggtcgagc
     7201 cacgcgccta tgcaccgcag ggattggccg aaaagctggg aatctcaaag cgcgttgaga
     7261 cgccggaagc cgtggccgac cggctgacaa aagcggttcg gcaggggtat gagcctgccc
     7321 tacaggccgc cgcaggagcg cgtgagatgc gcaagaaggc cgatcaagcc caagagacgg
     7381 cccgag
//
