LOCUS       Exported                6232 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 6232)
  AUTHORS   EJPARKER
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6232
                     /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..1434
                     /label=71_FWD_Gent
     promoter        1474..1502
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     1474..1502
                     /label=pmre116.for
     primer_bind     1523..1545
                     /label=FWD_Tri
     CDS             1691..2224
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(1691..1708)
                     /label=pAG-GmR-rev
     primer_bind     complement(1710..1723)
                     /label=pAG-GmR-rev
     primer_bind     2201..2218
                     /label=pAG408_rainbowFOR
     primer_bind     complement(2205..2237)
                     /label=72_RV_Gent
     primer_bind     complement(2205..2224)
                     /label=Gent.rev
     primer_bind     complement(2205..2224)
                     /label=pmre113.for
     primer_bind     complement(2207..2224)
                     /label=NotI_GmR_rev
     primer_bind     2225..2240
                     /label=FWD_primer_PnptII
     promoter        2231..2567
                     /locus_tag=""
                     /label=p-nptII promoter
                     /label=p-nptII
     primer_bind     2231..2250
                     /label=11x-nptII-FL-fw
     primer_bind     2231..2245
                     /label=lederm_nptII.fw
     primer_bind     2380..2404
                     /label=Fragment 2.FOR
     primer_bind     complement(2380..2404)
                     /label=Fragment 1.REV
     primer_bind     2381..2399
                     /label=pFru97xpMRE100_nptIIGib_rv
     primer_bind     complement(2566..2598)
                     /label=REV_primer_PnptII_mTq2/sG2
     primer_bind     complement(2566..2595)
                     /label=REV_primer_PnptII_mCl/mO/mSc/sY2
     primer_bind     complement(2566..2591)
                     /label=REV_primer_PnptII_mTagBFP2
     primer_bind     2584..2601
                     /label=XhoI_Ptac_XFP_for
     CDS             2586..3281
                     /codon_start=1
                     /locus_tag=""
                     /product=""
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     2586..2603
                     /label=XFPs for
     primer_bind     complement(2586..2603)
                     /label=Ptuf rev
     primer_bind     complement(3262..3301)
                     /label=REV_primer_mTq2
     primer_bind     complement(3263..3285)
                     /label=NotI_XFP_rev
     primer_bind     complement(3263..3284)
                     /label=nextgenFP.rv
     primer_bind     complement(3265..3301)
                     /label=REV_primer_mO
     primer_bind     complement(3265..3284)
                     /label=11x-FL-rv
     primer_bind     complement(3266..3301)
                     /label=REV_primer_sY2
     primer_bind     complement(3266..3284)
                     /label=REV_sYFP2
     primer_bind     complement(3266..3284)
                     /label=Ptac+eXFP_rev
     primer_bind     complement(3267..3284)
                     /label=XFPs rev KpnI
     primer_bind     3282..3296
                     /label=Gib_Gent.rev
     primer_bind     complement(3289..3308)
                     /label=FWD_pFru_ins
     primer_bind     3292..3314
                     /label=Gib_Gent.for.cor
     terminator      3298..3392
                     /locus_tag=""
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3436..4095
                     /codon_start=1
                     /locus_tag=""
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3497..3525)
                     /label=pMRE130inserts_rev
     primer_bind     complement(4080..4095)
                     /label=Chloramp_rev
     terminator      4160..4246
                     /locus_tag=""
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB ge
                     ne"
     primer_bind     complement(4251..4268)
                     /label=Gib_Gent.for
     primer_bind     complement(4256..4295)
                     /label=Fragment 2.REV
     primer_bind     complement(4256..4273)
                     /label=pMRE130_cassette2_RV
     primer_bind     complement(4257..4273)
                     /label=cassette_pAG408overhang_RV
     primer_bind     4274..4300
                     /label=pHimarEm1+XmaI_FWD
     repeat_region   4363..4391
                     /label=IR2
     primer_bind     4637..4653
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4733..4744)
                     /label=cassette_pAG408overhang_RV
     oriT            4740..4848
                     /note="incP origin of transfer"
     protein_bind    4995..5016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5031..5061
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5031..5060
                     /gene="lac prom"
                     /label=lac prom promoter
     protein_bind    5069..5085
                     /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    5105..6151
                     /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 agctttagta cctagattta gatgtctaaa
     1441 aagcggggta ccgaggacgc gtcaattctc gaattgacat aagcctgttc ggttcgtaaa
     1501 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     1561 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct
     1621 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     1681 agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1741 aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt
     1801 caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta
     1861 ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat
     1921 cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc
     1981 caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca
     2041 ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct
     2101 tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta
     2161 tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac
     2221 ctaaagctcg cacgctgccg caagcactca gggcgcaagg gctgctaaag gaagcggaac
     2281 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     2341 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     2401 tggcgatagc tagactgggc ggtcttatgg acagcaagcg aaccggaatt gccagctggg
     2461 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     2521 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatacta gtggaggaag
     2581 aaaaaatggt gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
     2641 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2701 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct
     2761 gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcatcaag caccccgccg
     2821 acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     2881 acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc
     2941 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     3001 agaagacaat gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga
     3061 agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca
     3121 agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca
     3181 agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     3241 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaattagct gagcttggac
     3301 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     3361 gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga
     3421 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     3481 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     3541 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     3601 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg
     3661 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     3721 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     3781 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     3841 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     3901 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     3961 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc
     4021 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     4081 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     4141 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4201 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgctaggagc
     4261 ttgcggccgc cagcccgggc aattcgaggg gtatcgctct tgaagggaac tatgttgaat
     4321 aataaaaacg aattttgaca aaaaaatgtg tttttctttg ttagaccggg gacttatcag
     4381 ccaacctgtt atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4561 ctgcggccaa cttacttctg acaacgatcg aggcgattaa gttgggtaac gccagggttt
     4621 tcccagtcac gacgttgtaa aacgacggcc agtgaattaa ttcttgaaga cgaaagggcc
     4681 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagagcttac
     4741 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     4801 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     4861 tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     4921 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcgggt
     4981 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5041 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5101 acatatggaa aaaaaggaat ttcgtgtttt gataaaatac tgttttctga agggaaaaaa
     5161 tacggtggaa gcaaaaactt ggcttgataa tgagtttccg gactctgccc cagggaaatc
     5221 aacaataatt gattggtatg caaaattcaa gcgtggtgaa atgagcacgg aggacggtga
     5281 acgcagtgga cgcccgaaag aggtggttac cgacgaaaac atcaaaaaaa tccacaaaat
     5341 gattttgaat gaccgtaaaa tgaagttgat cgagatagca gaggccttaa agatatcaaa
     5401 ggaacgtgtt ggtcatatca ttcatcaata tttggatatg cggaagctct gtgcaaaatg
     5461 ggtgccgcgc gagctcacat ttgaccaaaa acaacaacgt gttgatgatt ctgagcggtg
     5521 tttgcagctg ttaactcgta atacacccga gtttttccgt cgatatgtga caatggatga
     5581 aacatggctc catcactaca ctcctgagtc caatcgacag tcggctgagt ggacagcgac
     5641 cggtgaaccg tctccgaagc gtggaaagac tcaaaagtcc gctggcaaag taatggcctc
     5701 tgttttttgg gatgcgcatg gaataatttt tatcgattat cttgagaagg gaaaaaccat
     5761 caacagtgac tattatatgg cgttattgga gcgtttgaag gtcgaaatcg cggcaaaacg
     5821 gccccacatg aagaagaaaa aagtgttgtt ccaccaagac aacgcaccgt gccacaagtc
     5881 attgagaacg atggcaaaaa ttcatgaatt gggcttcgaa ttgcttcccc acccaccgta
     5941 ttctccagat ctggccccca gcgacttttt cttgttctca gacctcaaaa ggatgctcgc
     6001 agggaaaaaa tttggctgca atgaagaggt gatcgccgaa actgaggcct attttgaggc
     6061 aaaaccgaag gagtactacc aaaatggtat caaaaaattg gaaggtcgtt ataatcgttg
     6121 tatcgctctt gaagggaact atgttgaata ataaaaacga attttgacaa aaaaatgtgt
     6181 ttttctttgt tagaccggaa tctgcagaat tccagcacac tggcggccgt ta
//
