LOCUS       Exported                6272 bp DNA     circular SYN 23-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6272)
  AUTHORS   EJPARKER
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   274..302
                     /label=IR1
     rep_origin      587..975
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     complement(1350..1377)
                     /label=pHimarEm1+XbaI_RV2
     primer_bind     1357..1399
                     /label=Fragment 1.FOR
     primer_bind     1380..1405
                     /locus_tag=""
                     /label=Geneious name: MCS primer
                     /note="Geneious name: MCS primer"
     primer_bind     1380..1405
                     /label=pFru97 MCS fw
     primer_bind     1380..1403
                     /label=cassette_pAG408overhang_FWD
     primer_bind     1380..1400
                     /label=REV_pFru_ins
     primer_bind     1380..1399
                     /label=cassette5_pGRG36overhang_FWD
     primer_bind     1398..1433
                     /label=73_FWD_Kan
     primer_bind     1417..1453
                     /label=pMRE12-6/7/8/9for
     primer_bind     1417..1453
                     /label=pMRE123for
     CDS             1470..2264
                     /codon_start=1
                     /label=Kanamycin resistance
                     /note="Length: 795;Found at strand: positive;Start codon: 
                     ATG"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(2218..2264)
                     /label=pMRE12-6/7/8/9rev
     primer_bind     complement(2245..2277)
                     /label=74_RV_Kan
     primer_bind     2264..2280
                     /label=FWD_primer_PnptII
     promoter        2271..2607
                     /locus_tag=""
                     /label=p-nptII promoter
                     /label=p-nptII
     primer_bind     2271..2290
                     /label=11x-nptII-FL-fw
     primer_bind     2271..2285
                     /label=lederm_nptII.fw
     primer_bind     2420..2444
                     /label=Fragment 2.FOR
     primer_bind     complement(2420..2444)
                     /label=Fragment 1.REV
     primer_bind     complement(2420..2444)
                     /label=pntpII_1_REV
     primer_bind     2420..2444
                     /label=pntpII_1_FWD
     primer_bind     2421..2439
                     /label=pFru97xpMRE100_nptIIGib_rv
     primer_bind     complement(2606..2638)
                     /label=REV_primer_PnptII_mTq2/sG2
     primer_bind     complement(2606..2635)
                     /label=REV_primer_PnptII_mCl/mO/mSc/sY2
     primer_bind     complement(2606..2631)
                     /label=REV_primer_PnptII_mTagBFP2
     primer_bind     2624..2641
                     /label=XhoI_Ptac_XFP_for
     CDS             2626..3321
                     /codon_start=1
                     /locus_tag=""
                     /product=""
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     2626..2643
                     /label=XFPs for
     primer_bind     complement(2626..2643)
                     /label=Ptuf rev
     primer_bind     complement(3302..3341)
                     /label=REV_primer_mTq2
     primer_bind     complement(3303..3325)
                     /label=NotI_XFP_rev
     primer_bind     complement(3303..3324)
                     /label=nextgenFP.rv
     primer_bind     complement(3305..3341)
                     /label=REV_primer_mO
     primer_bind     complement(3305..3324)
                     /label=11x-FL-rv
     primer_bind     complement(3306..3341)
                     /label=REV_primer_sY2
     primer_bind     complement(3306..3324)
                     /label=REV_sYFP2
     primer_bind     complement(3306..3324)
                     /label=Ptac+eXFP_rev
     primer_bind     complement(3307..3324)
                     /label=XFPs rev KpnI
     primer_bind     3322..3336
                     /label=Gib_Gent.rev
     primer_bind     complement(3329..3348)
                     /label=FWD_pFru_ins
     primer_bind     3332..3354
                     /label=Gib_Gent.for.cor
     terminator      3338..3432
                     /locus_tag=""
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3476..4135
                     /codon_start=1
                     /locus_tag=""
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3537..3565)
                     /label=pMRE130inserts_rev
     primer_bind     complement(4120..4135)
                     /label=Chloramp_rev
     terminator      4200..4286
                     /locus_tag=""
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB ge
                     ne"
     primer_bind     complement(4291..4308)
                     /label=Gib_Gent.for
     primer_bind     complement(4296..4335)
                     /label=Fragment 2.REV
     primer_bind     complement(4296..4313)
                     /label=pMRE130_cassette2_RV
     primer_bind     complement(4297..4313)
                     /label=cassette_pAG408overhang_RV
     primer_bind     4314..4340
                     /label=pHimarEm1+XmaI_FWD
     repeat_region   4403..4431
                     /label=IR2
     primer_bind     4677..4693
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4773..4784)
                     /label=cassette_pAG408overhang_RV
     oriT            4780..4888
                     /note="incP origin of transfer"
     protein_bind    5035..5056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5071..5101
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5071..5100
                     /gene="lac prom"
                     /label=lac prom promoter
     protein_bind    5109..5125
                     /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)."
     misc_feature    5145..6191
                     /label=Transposase
                     /label=nonstandard type:  ORF
                     /note="/sequence=ORF_2 rf(1)"
ORIGIN
        1 ctagatgcat gctcgagcgg ccgccagtgt gatggatttg aaaaaggaag agtatgagta
       61 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      241 gttttccaat gatgagcact tttaaagttc tgctaacagg ttggctgata agtccccggt
      301 ctgacacata gatggcgtcg ctagtattaa atgcatatta tttttatata gtaccaacct
      361 tcaaatgatt cccagatctc aaactggaac aacactcaac cctatctcgg tctattcttt
      421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      541 ggaaatgtgc gcggaaccag atccccacga tgcgtccggc gtagaggatc tgaagatcag
      601 cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg
      661 tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac gtactaagct
      721 ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg tttgaacaat
      781 aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt ataagaaaaa
      841 aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca acaagccagg
      901 gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttcagtg acacaggaac
      961 acttaacggc tgacagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1021 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggttaatt
     1081 aagggctgca ggaattcgat atcaagctta tcgataccgt cgaaattctg attaataatt
     1141 tgtttaaatt tttcgtttgg cgtgaggtat ccaagtcttt tacgaggtcg attattgagt
     1201 ttattttcaa tccacttaat ctgtttgttg gttacttcac taaagtcctt accctttggg
     1261 atatactgcc tgataagccc gttggtgttt tcattggcac cacgttccca tgagtggtat
     1321 ggtttgcaaa aatagaattt tatttccaat ttttgcgcaa tttcctcgtg ctctagaata
     1381 taaactgcca ggaattgggg atcggattcg agctaagaat tcccttgggg tatctttaaa
     1441 tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca ccggaattga
     1501 aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct cctgctaagg
     1561 tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac agccggtata
     1621 aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg gaaggaaagc
     1681 tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat ctgctcatga
     1741 gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tggacaaagc cctgaaaaga
     1801 ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata tcggattgtc
     1861 cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg aataacgatc
     1921 tggccgatgt ggattgcgaa aactgggaag aagacactcc atttaaagat ccgcgcgagc
     1981 tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc cacggcgacc
     2041 tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt gatcttggga
     2101 gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg atcagggagg
     2161 atgtcgggga agaacagtat gtcgagctat tttttgactt actggggatc aagcctgatt
     2221 gggagaaaat aaaatattat attttactgg atgaattgtt ttagagctcg cacgctgccg
     2281 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
     2341 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     2401 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     2461 ggtcttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     2521 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     2581 atcaagatct gatcaagaga caggatacta gtggaggaag aaaaaatggt gagcaagggc
     2641 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc
     2701 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2761 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag
     2821 ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga ctactataag
     2881 cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc
     2941 gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc
     3001 cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa
     3061 gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc
     3121 ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag
     3181 aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac
     3241 aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc
     3301 ggcatggacg agctgtacaa gtaattagct gagcttggac tcctgttgat agatccagta
     3361 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     3421 attggtgaga atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga
     3481 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     3541 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     3601 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     3661 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     3721 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     3781 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     3841 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     3901 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     3961 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     4021 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg
     4081 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     4141 ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc
     4201 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     4261 tgaacgctct cctgagtagg acaaatccgc cgctaggagc ttgcggccgc cagcccgggc
     4321 aattcgaggg gtatcgctct tgaagggaac tatgttgaat aataaaaacg aattttgaca
     4381 aaaaaatgtg tttttctttg ttagaccggg gacttatcag ccaacctgtt atgtggcgcg
     4441 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4501 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4561 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4621 acaacgatcg aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     4681 aacgacggcc agtgaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     4741 taggttaatg tcatgataat aatggtttct tagagcttac ggccagcctc gcagagcagg
     4801 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     4861 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     4921 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     4981 aatcgctata atgaccccga agcagggtta tgcagcgggt gagcgcaacg caattaatgt
     5041 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     5101 gtgtggaatt gtgagcggat aacaatttca cacaggaaac acatatggaa aaaaaggaat
     5161 ttcgtgtttt gataaaatac tgttttctga agggaaaaaa tacggtggaa gcaaaaactt
     5221 ggcttgataa tgagtttccg gactctgccc cagggaaatc aacaataatt gattggtatg
     5281 caaaattcaa gcgtggtgaa atgagcacgg aggacggtga acgcagtgga cgcccgaaag
     5341 aggtggttac cgacgaaaac atcaaaaaaa tccacaaaat gattttgaat gaccgtaaaa
     5401 tgaagttgat cgagatagca gaggccttaa agatatcaaa ggaacgtgtt ggtcatatca
     5461 ttcatcaata tttggatatg cggaagctct gtgcaaaatg ggtgccgcgc gagctcacat
     5521 ttgaccaaaa acaacaacgt gttgatgatt ctgagcggtg tttgcagctg ttaactcgta
     5581 atacacccga gtttttccgt cgatatgtga caatggatga aacatggctc catcactaca
     5641 ctcctgagtc caatcgacag tcggctgagt ggacagcgac cggtgaaccg tctccgaagc
     5701 gtggaaagac tcaaaagtcc gctggcaaag taatggcctc tgttttttgg gatgcgcatg
     5761 gaataatttt tatcgattat cttgagaagg gaaaaaccat caacagtgac tattatatgg
     5821 cgttattgga gcgtttgaag gtcgaaatcg cggcaaaacg gccccacatg aagaagaaaa
     5881 aagtgttgtt ccaccaagac aacgcaccgt gccacaagtc attgagaacg atggcaaaaa
     5941 ttcatgaatt gggcttcgaa ttgcttcccc acccaccgta ttctccagat ctggccccca
     6001 gcgacttttt cttgttctca gacctcaaaa ggatgctcgc agggaaaaaa tttggctgca
     6061 atgaagaggt gatcgccgaa actgaggcct attttgaggc aaaaccgaag gagtactacc
     6121 aaaatggtat caaaaaattg gaaggtcgtt ataatcgttg tatcgctctt gaagggaact
     6181 atgttgaata ataaaaacga attttgacaa aaaaatgtgt ttttctttgt tagaccggaa
     6241 tctgcagaat tccagcacac tggcggccgt ta
//
