LOCUS       Exported                5436 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 5436)
  AUTHORS   EJPARKER
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5436
                     /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.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     1390..1426
                     /label=FWD_primer_PnptII
     promoter        1417..1752
                     /locus_tag=""
                     /label=p-nptII promoter
                     /label=p-nptII
     primer_bind     1566..1584
                     /label=pFru97xpMRE100_nptIIGib_rv
     primer_bind     complement(1751..1781)
                     /label=REV_primer_PnptII_mTq2/sG2
     primer_bind     complement(1751..1771)
                     /label=REV_primer_PnptII_mCl/mO/mSc/sY2
     primer_bind     complement(1751..1771)
                     /label=REV_primer_PnptII_mTagBFP2
     primer_bind     1761..1791
                     /label=FWD_primer_mTq2/sGFP2
     primer_bind     1761..1788
                     /label=FWD_primer_mCl/mO/mSc/sY2
     CDS             1773..2485
                     /codon_start=1
                     /locus_tag=""
                     /product=""
                     /label=mTurquoise2
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     1773..1788
                     /label=FWD_sYFP2
     primer_bind     1773..1786
                     /label=XFPs for
     primer_bind     complement(2290..2310)
                     /label=FWD_primer_PnptII
     primer_bind     complement(2436..2505)
                     /label=REV_primer_sG2
     primer_bind     complement(2466..2505)
                     /label=REV_primer_mTq2
     primer_bind     complement(2467..2489)
                     /label=NotI_XFP_rev
     primer_bind     complement(2467..2488)
                     /label=nextgenFP.rv
     primer_bind     complement(2469..2505)
                     /label=REV_primer_mO
     primer_bind     complement(2469..2488)
                     /label=11x-FL-rv
     primer_bind     complement(2470..2505)
                     /label=REV_primer_sY2
     primer_bind     complement(2470..2488)
                     /label=REV_sYFP2
     primer_bind     complement(2470..2488)
                     /label=Ptac+eXFP_rev
     primer_bind     complement(2471..2488)
                     /label=XFPs rev KpnI
     primer_bind     2486..2500
                     /label=Gib_Gent.rev
     primer_bind     complement(2493..2512)
                     /label=FWD_pFru_ins
     primer_bind     2496..2518
                     /label=Gib_Gent.for.cor
     terminator      2502..2596
                     /locus_tag=""
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2640..3299
                     /codon_start=1
                     /locus_tag=""
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2701..2729)
                     /label=pMRE130inserts_rev
     primer_bind     complement(3284..3299)
                     /label=Chloramp_rev
     terminator      3364..3450
                     /locus_tag=""
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB ge
                     ne"
     primer_bind     complement(3455..3472)
                     /label=Gib_Gent.for
     primer_bind     complement(3460..3499)
                     /label=Fragment.REV
     primer_bind     complement(3460..3477)
                     /label=pMRE130_cassette2_RV
     primer_bind     complement(3461..3477)
                     /label=cassette_pAG408overhang_RV
     primer_bind     3478..3504
                     /label=pHimarEm1+XmaI_FWD
     repeat_region   3567..3595
                     /label=IR2
     primer_bind     3841..3857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3937..3948)
                     /label=cassette_pAG408overhang_RV
     oriT            3944..4052
                     /note="incP origin of transfer"
     protein_bind    4199..4220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4235..4265
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        4235..4264
                     /gene="lac prom"
                     /label=lac prom promoter
     protein_bind    4273..4289
                     /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    4309..5355
                     /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 agctcgcacg ctgccgcagc actcagggcg
     1441 caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc
     1501 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa
     1561 gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc
     1621 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     1681 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca
     1741 agagacagga tactagtgga ggaagaaaaa aatgagcaag ggcgaggagc tgttcaccgg
     1801 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1861 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1921 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgtcctggg gcgtgcagtg
     1981 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2041 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2101 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2161 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacttta gcgacaacgt
     2221 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     2281 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2341 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     2401 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2461 tctcggcatg gacgagctgt acaagtaatt agctgagctt ggactcctgt tgatagatcc
     2521 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     2581 ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa
     2641 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     2701 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     2761 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     2821 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     2881 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     2941 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     3001 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     3061 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     3121 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     3181 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg
     3241 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     3301 ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg
     3361 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3421 tcggtgaacg ctctcctgag taggacaaat ccgccgctag gagcttgcgg ccgccagccc
     3481 gggcaattcg aggggtatcg ctcttgaagg gaactatgtt gaataataaa aacgaatttt
     3541 gacaaaaaaa tgtgtttttc tttgttagac cggggactta tcagccaacc tgttatgtgg
     3601 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3661 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3721 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3781 tctgacaacg atcgaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     3841 gtaaaacgac ggccagtgaa ttaattcttg aagacgaaag ggcctcgtga tacgcctatt
     3901 tttataggtt aatgtcatga taataatggt ttcttagagc ttacggccag cctcgcagag
     3961 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     4021 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     4081 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     4141 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc gggtgagcgc aacgcaatta
     4201 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     4261 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacacatat ggaaaaaaag
     4321 gaatttcgtg ttttgataaa atactgtttt ctgaagggaa aaaatacggt ggaagcaaaa
     4381 acttggcttg ataatgagtt tccggactct gccccaggga aatcaacaat aattgattgg
     4441 tatgcaaaat tcaagcgtgg tgaaatgagc acggaggacg gtgaacgcag tggacgcccg
     4501 aaagaggtgg ttaccgacga aaacatcaaa aaaatccaca aaatgatttt gaatgaccgt
     4561 aaaatgaagt tgatcgagat agcagaggcc ttaaagatat caaaggaacg tgttggtcat
     4621 atcattcatc aatatttgga tatgcggaag ctctgtgcaa aatgggtgcc gcgcgagctc
     4681 acatttgacc aaaaacaaca acgtgttgat gattctgagc ggtgtttgca gctgttaact
     4741 cgtaatacac ccgagttttt ccgtcgatat gtgacaatgg atgaaacatg gctccatcac
     4801 tacactcctg agtccaatcg acagtcggct gagtggacag cgaccggtga accgtctccg
     4861 aagcgtggaa agactcaaaa gtccgctggc aaagtaatgg cctctgtttt ttgggatgcg
     4921 catggaataa tttttatcga ttatcttgag aagggaaaaa ccatcaacag tgactattat
     4981 atggcgttat tggagcgttt gaaggtcgaa atcgcggcaa aacggcccca catgaagaag
     5041 aaaaaagtgt tgttccacca agacaacgca ccgtgccaca agtcattgag aacgatggca
     5101 aaaattcatg aattgggctt cgaattgctt ccccacccac cgtattctcc agatctggcc
     5161 cccagcgact ttttcttgtt ctcagacctc aaaaggatgc tcgcagggaa aaaatttggc
     5221 tgcaatgaag aggtgatcgc cgaaactgag gcctattttg aggcaaaacc gaaggagtac
     5281 taccaaaatg gtatcaaaaa attggaaggt cgttataatc gttgtatcgc tcttgaaggg
     5341 aactatgttg aataataaaa acgaattttg acaaaaaaat gtgtttttct ttgttagacc
     5401 ggaatctgca gaattccagc acactggcgg ccgtta
//
