LOCUS       Exported                7124 bp DNA     circular SYN 22-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7124)
  AUTHORS   EJPARKER
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7124
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     complement(22..37)
                     /label=tn5_insert_REV
     repeat_region   274..302
                     /label=IR1
     primer_bind     complement(373..393)
                     /label=pHimar_gibson_RV
     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     1029..1050
                     /label=pMRE130_pTE_tetAtetR_RV
     primer_bind     1054..1075
                     /label=test himar fwd
     primer_bind     complement(1641..1650)
                     /label=FWD_sYFP2
     primer_bind     complement(1641..1650)
                     /label=FWD_primer_mCl/mO/mSc/sY2
     primer_bind     1978..2005
                     /label=promErmF_pHimar_overhang_FWD
     primer_bind     complement(2254..2277)
                     /label=promErmF_pHimar_overhang_RVa
     CDS             2278..3078
                     /codon_start=1
                     /label=Erythromycin resistance ErmF
                     /label=nonstandard type:  ORF
                     /note="/sequence=ORF_1 rf(1)"
                     /translation="MTKKKLPVRFTGQHFTIDKVLIKDAIRQANISNQDTVLDIGAGKG
                     FLTVHLLKIANNVVAIENDTALVEHLRKLFSDARNVQVVGCDFRNFAVPKFPFKVVSNI
                     PYGITSDIFKILMFESLGNFLGGSIVLQLEPTQKLFSRKLYNPYTVFYHTFFDLKLVYE
                     VGPESFLPPPTVKSALLNIKRKHLFFDFKFKAKYLAFISYLLEKPDLSVKTALKSIFRK
                     SQVRSISEKFGLNLNAQIVCLSPSQWLNCFLEMLEVVPEKFHPS"
     primer_bind     complement(3054..3084)
                     /label=Vector.REV
     primer_bind     complement(3054..3084)
                     /label=pHimarEm1_EmR+XbaI_RV
     primer_bind     complement(3054..3078)
                     /label=pHimarEm1_EmR+XmaI_RV
     primer_bind     3064..3108
                     /label=Fragment 1.FOR
     primer_bind     3085..3109
                     /label=cassette_pAG408overhang_FWD
     primer_bind     3086..3111
                     /locus_tag=""
                     /label=Geneious name: MCS primer
                     /note="Geneious name: MCS primer"
     primer_bind     3086..3111
                     /label=pFru97 MCS fw
     primer_bind     3086..3106
                     /label=REV_pFru_ins
     primer_bind     3086..3105
                     /label=cassette5_pGRG36overhang_FWD
     primer_bind     3096..3132
                     /label=FWD_primer_PnptII
     promoter        3123..3459
                     /locus_tag=""
                     /label=p-nptII promoter
                     /label=p-nptII
     primer_bind     3123..3142
                     /label=11x-nptII-FL-fw
     primer_bind     3123..3137
                     /label=lederm_nptII.fw
     primer_bind     3272..3296
                     /label=Fragment 2.FOR
     primer_bind     complement(3272..3296)
                     /label=pntpII_1_REV
     primer_bind     3272..3296
                     /label=pntpII_1_FWD
     primer_bind     complement(3272..3296)
                     /label=Fragment 1.REV
     primer_bind     3273..3291
                     /label=pFru97xpMRE100_nptIIGib_rv
     primer_bind     complement(3394..3414)
                     /label=pntpII_2_REV
     primer_bind     3394..3414
                     /label=pntpII_2_FWD
     primer_bind     complement(3438..3472)
                     /label=rbs_1_REV
     primer_bind     3438..3472
                     /label=rbs_1_FWD
     primer_bind     complement(3458..3490)
                     /label=REV_primer_PnptII
     primer_bind     complement(3458..3487)
                     /label=REV_primer_PnptII (3)
     primer_bind     complement(3458..3483)
                     /label=REV_primer_PnptII (2)
     primer_bind     3476..3493
                     /label=XhoI_Ptac_XFP_for
     CDS             3478..4173
                     /codon_start=1
                     /locus_tag=""
                     /product=""
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     3478..3495
                     /label=XFPs for
     primer_bind     complement(3478..3495)
                     /label=Ptuf rev
     primer_bind     complement(4154..4193)
                     /label=REV_primer_mTq2
     primer_bind     complement(4155..4177)
                     /label=NotI_XFP_rev
     primer_bind     complement(4155..4176)
                     /label=nextgenFP.rv
     primer_bind     complement(4157..4193)
                     /label=REV_primer_mO
     primer_bind     complement(4157..4176)
                     /label=11x-FL-rv
     primer_bind     complement(4158..4193)
                     /label=REV_primer_sY2
     primer_bind     complement(4158..4176)
                     /label=REV_sYFP2
     primer_bind     complement(4158..4176)
                     /label=Ptac+eXFP_rev
     primer_bind     complement(4159..4176)
                     /label=XFPs rev KpnI
     primer_bind     4174..4188
                     /label=Gib_Gent.rev
     primer_bind     complement(4181..4200)
                     /label=FWD_pFru_ins
     primer_bind     4184..4206
                     /label=Gib_Gent.for.cor
     terminator      4190..4284
                     /locus_tag=""
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4328..4987
                     /codon_start=1
                     /locus_tag=""
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4389..4417)
                     /label=pMRE130inserts_rev
     primer_bind     complement(4972..4987)
                     /label=Chloramp_rev
     primer_bind     5022..5042
                     /label=ARB-RB-PCR1
     terminator      5052..5138
                     /locus_tag=""
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB ge
                     ne"
     primer_bind     5124..5144
                     /label=ARB-RB-PCR2
     primer_bind     complement(5143..5160)
                     /label=Gib_Gent.for
     primer_bind     complement(5148..5187)
                     /label=Fragment 2.REV
     primer_bind     complement(5148..5165)
                     /label=cassette_pGRG36overhang_RV
     primer_bind     complement(5148..5165)
                     /label=pMRE130_cassette2_RV
     primer_bind     complement(5149..5165)
                     /label=cassette_pAG408overhang_RV
     primer_bind     5166..5192
                     /label=Vector.FOR
     primer_bind     5172..5192
                     /label=pHimarEm1+XbaI_FWD
     primer_bind     5242..5263
                     /label=seq1_pHimarv2
     repeat_region   5255..5283
                     /label=IR2
     primer_bind     complement(5380..5408)
                     /label=test himar v2 rvs
     primer_bind     complement(5461..5482)
                     /label=pHimar_gibson2_RV
     primer_bind     5477..5496
                     /label=pHimar_gibson2_FWD
     primer_bind     5513..5536
                     /label=pMRE130_cassette2_FWD
     primer_bind     5513..5536
                     /label=pMRE130_fwd_new
     primer_bind     5515..5530
                     /label=tn5_insert_FWD
     primer_bind     5529..5545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5529..5545
                     /label=M13F
     primer_bind     5603..5624
                     /label=pMRE130_pTE_tetAtetR_RV
     primer_bind     complement(5625..5636)
                     /label=cassette_pAG408overhang_RV
     oriT            5632..5740
                     /note="incP origin of transfer"
     protein_bind    5887..5908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5923..5953
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5923..5952
                     /gene="lac prom"
                     /label=lac prom promoter
     protein_bind    5961..5977
                     /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    5997..7043
                     /label=Transposase
                     /label=nonstandard type:  ORF
                     /note="/sequence=ORF_2 rf(1)"
     primer_bind     complement(6041..6060)
                     /label=seq2_pHimarv2
     primer_bind     7094..7117
                     /label=tn5_insert_REV
     primer_bind     join(7114..7124,1..18)
                     /label=pTuf fwd NotI
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 ctttgcaaac
     1381 tcctttccat tgtcagccgt aattgtgtgt attaagtttt tcactttccg cagtgcccat
     1441 actgcaatct tagctaccgg gatggcttct tttcccgaca acttgcgtat ccagaccctg
     1501 cttgttgctc tgtcgttaat ggtaagaatg gcacctttgt ggttcttacc aataattgta
     1561 tctatctcta aatcaccaaa tctctccttc agttccacta tctcgggacg ctcatcaata
     1621 tccaccctgc ctgggataaa tcctcgccct gcatttttag aaccacgttt ggcatacctg
     1681 cgaccttgtc tgcgaagata tttgtgcagt ttgccacccc gccgcttatc ctcccaaatc
     1741 cagcgatata tcgtttcgtg agataccatc gcaattccct ccaagcggct cctgccgaca
     1801 atctgctccg ggctgaatcc tttcttcaac agctttatta tccgttttct cattgccggt
     1861 gtaagcactt ccttgcgatg tttttgctgc ttgcgcctgt ctgcttttcg ctgggcaagc
     1921 tccatgctat agctaccact tcgggcgtcg caattgcgct ttatctccct gtaaacagtg
     1981 cttttatcta ctccgatagc ttccgctatt gcttttttgc tcatcggtat ttgcaacatc
     2041 atagaaattg catacctttg ttcctcggtt atatgtttgc tcatctgcaa cttttttttc
     2101 tttggacgga caattaaagc aaagatagca aactttatcc attcagagtg agagaaaggg
     2161 ggacattgtc tctctttcct ctctgaaaaa taaatgtttt tattgcttat tatccgcacc
     2221 caaaaagttg catttataag ttgaactcaa gaagtattca cctgtaagaa gttactaatg
     2281 acaaaaaaga aattgcccgt tcgttttacg ggtcagcact ttactattga taaagtgcta
     2341 ataaaagatg caataagaca agcaaatata agtaatcagg atacggtttt agatattggg
     2401 gcaggcaagg ggtttcttac tgttcattta ttaaaaatcg ccaacaatgt tgttgctatt
     2461 gaaaacgaca cagctttggt tgaacattta cgaaaattat tttctgatgc ccgaaatgtt
     2521 caagttgtcg gttgtgattt taggaatttt gcagttccga aatttccttt caaagtggtg
     2581 tcaaatattc cttatggcat tacttccgat attttcaaaa tcctgatgtt tgagagtctt
     2641 ggaaattttc tgggaggttc cattgtcctt caattagaac ctacacaaaa gttattttcg
     2701 aggaagcttt acaatccata taccgttttc tatcatactt tttttgattt gaaacttgtc
     2761 tatgaggtag gtcctgaaag tttcttgcca ccgccaactg tcaaatcagc cctgttaaac
     2821 attaaaagaa aacacttatt ttttgatttt aagtttaaag ccaaatactt agcatttatt
     2881 tcctatctgt tagagaaacc tgatttatct gtaaaaacag ctttaaagtc gattttcagg
     2941 aaaagtcagg tcaggtcaat ttcggaaaaa ttcggtttaa accttaatgc tcaaattgtt
     3001 tgtttgtctc caagtcaatg gttaaactgt tttttggaaa tgctggaagt tgtccctgaa
     3061 aaatttcatc cttcgtagtc tagacataaa ctgccaggaa ttggggatcg gattcgagct
     3121 cgcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     3181 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     3241 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     3301 gctagactgg gcggtcttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     3361 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     3421 atggcgcagg ggatcaagat ctgatcaaga gacaggatac tagtggagga agaaaaaatg
     3481 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc
     3541 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3601 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc
     3661 ctgtcccctc agttcatgta cggctccagg gccttcatca agcaccccgc cgacatcccc
     3721 gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3781 gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac
     3841 aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca
     3901 atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac
     3961 attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc
     4021 tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac
     4081 atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc
     4141 cactccaccg gcggcatgga cgagctgtac aagtaattag ctgagcttgg actcctgttg
     4201 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     4261 cgggcgtttt ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga
     4321 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     4381 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     4441 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     4501 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     4561 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     4621 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     4681 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     4741 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     4801 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     4861 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     4921 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     4981 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta
     5041 gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     5101 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgctagga gcttgcggcc
     5161 gccagcccgg gcaattcgag gggtatcgct cttgaaggga actatgttga ataataaaaa
     5221 cgaattttga caaaaaaatg tgtttttctt tgttagaccg gggacttatc agccaacctg
     5281 ttatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5461 aacttacttc tgacaacgat cgaggcgatt aagttgggta acgccagggt tttcccagtc
     5521 acgacgttgt aaaacgacgg ccagtgaatt aattcttgaa gacgaaaggg cctcgtgata
     5581 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagagctt acggccagcc
     5641 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     5701 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     5761 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
     5821 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg gtgagcgcaa
     5881 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     5941 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acacatatgg
     6001 aaaaaaagga atttcgtgtt ttgataaaat actgttttct gaagggaaaa aatacggtgg
     6061 aagcaaaaac ttggcttgat aatgagtttc cggactctgc cccagggaaa tcaacaataa
     6121 ttgattggta tgcaaaattc aagcgtggtg aaatgagcac ggaggacggt gaacgcagtg
     6181 gacgcccgaa agaggtggtt accgacgaaa acatcaaaaa aatccacaaa atgattttga
     6241 atgaccgtaa aatgaagttg atcgagatag cagaggcctt aaagatatca aaggaacgtg
     6301 ttggtcatat cattcatcaa tatttggata tgcggaagct ctgtgcaaaa tgggtgccgc
     6361 gcgagctcac atttgaccaa aaacaacaac gtgttgatga ttctgagcgg tgtttgcagc
     6421 tgttaactcg taatacaccc gagtttttcc gtcgatatgt gacaatggat gaaacatggc
     6481 tccatcacta cactcctgag tccaatcgac agtcggctga gtggacagcg accggtgaac
     6541 cgtctccgaa gcgtggaaag actcaaaagt ccgctggcaa agtaatggcc tctgtttttt
     6601 gggatgcgca tggaataatt tttatcgatt atcttgagaa gggaaaaacc atcaacagtg
     6661 actattatat ggcgttattg gagcgtttga aggtcgaaat cgcggcaaaa cggccccaca
     6721 tgaagaagaa aaaagtgttg ttccaccaag acaacgcacc gtgccacaag tcattgagaa
     6781 cgatggcaaa aattcatgaa ttgggcttcg aattgcttcc ccacccaccg tattctccag
     6841 atctggcccc cagcgacttt ttcttgttct cagacctcaa aaggatgctc gcagggaaaa
     6901 aatttggctg caatgaagag gtgatcgccg aaactgaggc ctattttgag gcaaaaccga
     6961 aggagtacta ccaaaatggt atcaaaaaat tggaaggtcg ttataatcgt tgtatcgctc
     7021 ttgaagggaa ctatgttgaa taataaaaac gaattttgac aaaaaaatgt gtttttcttt
     7081 gttagaccgg aatctgcaga attccagcac actggcggcc gtta
//
