LOCUS       Exported                7212 bp ds-DNA     circular SYN 13-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7212)
  AUTHORS   Lu Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 18, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7212
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    187..326
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(428..619)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1075..1100)
                     /label=vector_rev
     misc_feature    1100^1101
                     /label=VB
     misc_feature    1108..1116
                     /label=RBS
     CDS             1122..1388
                     /codon_start=1
                     /label=GvpB
                     /translation="MSIQKSTNSSSLAEVIDRILDKGIVIDAFARVSVVGIEILTIEAR
                     VVIASVDTWLRYAEAVGLLRDDVEENGLPERSNSSEGQPRFSI"
     primer_bind     1411..1436
                     /label=Bm1seq
     primer_bind     complement(1420..1435)
                     /label=gvpB_rev
     misc_feature    1435^1436
                     /label=BR
     primer_bind     1436..1458
                     /label=gvpR_fwd
     misc_feature    1449..1457
                     /label=RBS
     CDS             1464..1730
                     /codon_start=1
                     /label=GvpR
                     /translation="MEIKKIMQAVNDFFGEHVAPPHKITSVEATEDEGWRVIVEVIEER
                     EYMKKYAKDEMLGTYECFVNKEKEVISFKRLDVRYRSAIGIEA"
     primer_bind     complement(1721..1742)
                     /label=gvpR_rev
     misc_feature    1742^1743
                     /label=RN
     primer_bind     1743..1763
                     /label=gvpN_fwd
     misc_feature    1744..1752
                     /label=RBS
     CDS             1759..2685
                     /codon_start=1
                     /label=GvpN
                     /translation="MTVLTDKRKKGSGAFIQDDETKEVLSRALSYLKSGYSIHFTGPAG
                     GGKTSLARALAKKRKRPVMLMHGNHELNNKDLIGDFTGYTSKKVIDQYVRSVYKKDEQV
                     SENWQDGRLLEAVKNGYTLIYDEFTRSKPATNNIFLSILEEGVLPLYGVKMTDPFVRVH
                     PDFRVIFTSNPAEYAGVYDTQDALLDRLITMFIDYKDIDRETAILTEKTDVEEDEARTI
                     VTLVANVRNRSGDENSSGLSLRASLMIATLATQQDIPIDGSDEDFQTLCIDILHHPLTK
                     CLDEENAKSKAEKIILEECKNIDTEEK"
     primer_bind     1789..1814
                     /label=Bm1.1seqF
     misc_feature    1821
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     2191..2210
                     /label=Bm2seqF
     primer_bind     complement(2656..2685)
                     /label=gvpN_rev
     misc_feature    2685
                     /label=NF
     primer_bind     2686..2708
                     /label=gvpG_fwd
     CDS             2686..2700
                     /codon_start=1
                     /label=GvpF
                     /translation="VEEK"
     misc_feature    2686
                     /label=FG
     misc_feature    2690..2698
                     /label=RBS
     CDS             2702..2968
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKRNEES"
     primer_bind     2914..2941
                     /label=Bm4seqF
     primer_bind     complement(2932..2953)
                     /label=gvpG_rev
     misc_feature    2953^2954
                     /label=GL
     primer_bind     2954..2973
                     /label=gvpL_fwd
     misc_feature    2955..2963
                     /label=RBS
     CDS             2969..3778
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    3039
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3108
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3558
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3689..3715
                     /label=Bm5seqF
     primer_bind     complement(3762..3779)
                     /label=gvpL_rev
     misc_feature    3779^3780
                     /label=LS
     primer_bind     3780..3804
                     /label=gvpS_fwd
     misc_feature    3780..3788
                     /label=RBS
     CDS             3796..4083
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(4099..4118)
                     /label=gvpS_rev
     misc_feature    4118^4119
                     /label=SK
     primer_bind     4119..4134
                     /label=gvpK_fwd
     misc_feature    4140..4148
                     /label=RBS
     CDS             4155..4439
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4430..4445)
                     /label=gvpK_rev
     misc_feature    4445^4446
                     /label=KJ
     primer_bind     4446..4464
                     /label=gvpJ_fwd
     misc_feature    4449..4457
                     /label=RBS
     CDS             4461..4763
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    4586
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4592..4613
                     /label=Bm6seqF
     primer_bind     complement(4758..4779)
                     /label=gvpJ_rev
     misc_feature    4779
                     /label=JT
     primer_bind     4780..4800
                     /label=gvpU_fwd
     misc_feature    4780
                     /label=TU
     misc_feature    4782..4790
                     /label=RBS
     CDS             4798..5214
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5137..5161
                     /label=GvpUseqF
     misc_feature    5367^5368
                     /label=UV
     primer_bind     5368..5385
                     /label=vector_fwd
     terminator      5484..5531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5598..5689
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5679..5684
                     /label=Emopec_63.3%
     CDS             5690..6550
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6624..7212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aacgccagca acgcggccct ttttacggtt cctggccttt tgctggcctt ttgctcacat
       61 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      121 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      181 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
      241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      301 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
      361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
      481 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
      541 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
      601 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atggggtaat
      661 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
      721 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
      781 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttg gcaccatgca
      841 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagag
      901 atagcttagc gatgtgaaag aagctggaga aaactttcaa gaaaaagttt cttcttcaga
      961 cgataaagag aagtcaaaaa ataagcggaa aataaaaggc gtaaatcaga ttaaaaaagc
     1021 aacggatatt aaaagctcta ctaagattaa aagttctaat gatattaaat catctacaga
     1081 tttaaaaaca atgggatcat aagcgaaagg agaaatgaga tatgtctatt caaaaaagta
     1141 ctaatagttc aagtttagca gaagtcattg accgtatttt agataaagga attgttattg
     1201 atgcttttgc aagagtttct gttgtaggaa ttgaaatttt aacgattgaa gcgcgagtgg
     1261 ttattgccag tgttgataca tggttacgct atgcagaagc agtagggctt cttcgtgacg
     1321 acgtagaaga aaacggtctt cctgaacgtt caaattcaag tgaagggcag ccgcgtttta
     1381 gtatttaata tgaagtggta tggaaccttc cttttttgag ggtggttcca gcttttagat
     1441 ctaactattg gaggctacta aaaatggaaa ttaaaaaaat tatgcaagcc gtgaacgact
     1501 ttttcggtga acacgtagct cctcctcata aaattacctc ggtggaagct actgaagatg
     1561 aaggttggag agttattgtt gaagtcattg aagaacgaga atatatgaaa aaatacgcca
     1621 aagatgaaat gctcggaacg tacgagtgct ttgtaaataa agaaaaagaa gtcatttcat
     1681 tcaaacgact cgacgtcaga tatagaagcg ccattggcat tgaagcataa acgataagat
     1741 ggcaggagga acgtaaaaat gaccgtctta acagacaaaa ggaaaaaagg cagtggagct
     1801 tttatacaag atgacgagac aaaagaggtt ctttcaagag cgctgagcta tttaaaatcc
     1861 ggctattcca ttcattttac aggtcctgcc ggcggaggca aaacctcttt agcgcgagcg
     1921 cttgctaaaa agagaaagcg tcctgtaatg ctgatgcacg ggaatcacga gctcaacaac
     1981 aaagatttaa ttggcgattt tacgggatac acgagcaaaa aagtaatcga ccagtacgtt
     2041 cgttctgtct ataaaaaaga tgaacaggtg agtgaaaact ggcaggatgg ccgattgctt
     2101 gaagctgtaa aaaatggcta tacgctgatt tacgacgaat ttactcgttc taagcctgcg
     2161 acgaataata tctttctatc gatattagaa gaaggcgtgc tgccgctgta tggagtaaaa
     2221 atgaccgatc cttttgtgcg cgtgcatccc gatttccgcg tcatcttcac aagcaatcca
     2281 gctgagtatg ccggcgtata tgatacgcaa gatgcgcttc tcgacaggtt aattaccatg
     2341 tttattgatt ataaagacat cgacagagag acagcgattt taacggagaa aacggacgta
     2401 gaagaagatg aagcgcgcac aattgtaacg ctcgtagcaa acgtgcgaaa ccgctctgga
     2461 gacgaaaaca gcagcggact tagcctgcgg gcttcgctta tgatcgctac ccttgccacg
     2521 cagcaagaca ttcctatcga tggaagtgac gaagattttc aaacgttatg tatcgatatt
     2581 ttgcatcatc cgcttaccaa atgtttggat gaagaaaatg caaaaagcaa agccgaaaaa
     2641 atcattttag aagaatgtaa gaatatagac actgaagaaa agtaagtaga agagaaataa
     2701 cgtgcttcac aaattagtaa ccgcacccat taaccttgta gtgaaaatcg gcgaaaaagt
     2761 acaggaagaa gctgataaac agctatatga ccttccgacg attcagcaaa agctcattca
     2821 gcttcaaatg atgtttgagc ttggtgaaat tccagaagaa gcgtttcaag aaaaagaaga
     2881 tgaattgtta atgaggtacg aaattgcgaa acgcagagaa attgaacaat gggaagagct
     2941 aacacaaaaa agaaatgagg aatcctagat gggagaatta ctgtatttat acggtttaat
     3001 tccaacaaaa gaagcagcag ccatagagcc gtttccatct tataaggggt ttgacggaga
     3061 acattcactg tacccaattg cgtttgatca ggtgacggct gtagtttcta agctggatgc
     3121 tgacacctat tcagaaaaag tgattcaaga aaaaatggag caggatatga gctggctgca
     3181 ggaaaaagca tttcatcatc acgaaacggt agccgctttg tacgaagaat ttacgatcat
     3241 tccattaaaa ttttgcacca tttataaagg tgaagaaagt ctgcaggcag ctattgagat
     3301 taacaaagaa aagatagaga attcactgac gctgcttcaa ggaaatgaag agtggaatgt
     3361 gaaaatttac tgtgatgata cagagcttaa aaaaggaatc agcgaaacga atgaaagcgt
     3421 gaaagcgaaa aaacaagaaa ttagtcactt atcaccagga agacagtttt ttgaaaagaa
     3481 aaaaatagat cagctgattg aaaaagaatt agagcttcac aaaaacaaag tgtgtgaaga
     3541 gatacatgac aagctaaaag aattatcgct ttatgactct gttaaaaaga attggagcaa
     3601 agacgtaact ggcgcagctg aacagatggc gtggaacagc gtgtttcttc tcccgtctct
     3661 gcagattact aagttcgtaa acgaaataga agagcttcag caaaggcttg aaaataaagg
     3721 ctggaagttt gaagtgacgg gaccatggcc gccctatcat ttctcgagct ttgcgtaaag
     3781 tgaggaatta acattatgtc tcttaaacaa tccatggaga ataaagatat tgctcttatt
     3841 gatattttag atgtcatttt agataaagga gtcgccatta aaggagactt aatcatttcc
     3901 atagctggcg tcgatttagt gtatttggat ttgcgggtgc ttatttcttc ggttgaaacg
     3961 cttgtgcaag caaaagaagg aaatcacaaa ccaatcactt ctgaacaatt tgataaacaa
     4021 aaggaagagt taatggacgc gaccggccag ccgagcaaat ggaccaatcc gcttggctcc
     4081 tgacccttgc gagcagtcgt atgctttctc gaattccgag cggttaagcg tgacagtcca
     4141 ggaggaatta atggatgcaa ccggtcagcc aagcaaatgg acgaatccac ttggatcctg
     4201 atcaagctga acaaggctta gcgcagcttg tgatgacagt tattgagcta ttgaggcaaa
     4261 tcgttgaacg tcatgccatg aggcgggtgg agggtggaac gttgacggac gaacaaattg
     4321 aaaacttagg aattgcacta atgaacttag aagaaaaaat ggacgagttg aaagaggtgt
     4381 tcggtctgga tgcagaagat ttaaatattg atcttggacc gctaggcagc ctgctttaag
     4441 cggtcagtag gaggaacagt atggcagtcg aacataatat gcagtcaagt acgattgtag
     4501 atgtgctcga aaagattttg gataaaggag tcgttatagc gggggacatc accgtaggaa
     4561 ttgcagatgt cgagctatta acgataaaga tccgcttgat tgtggcttcg gttgataagg
     4621 caaaagaaat cggcatggac tggtgggaaa atgatccgta tctcagttca aaaggagcca
     4681 ataacaaagc gctcgaagaa gaaaataaaa tgctgcatga gcggttaaaa acgcttgaag
     4741 aaaaaataga aacgaaacgt taaaaactgt acgctactta aaggaggaaa gaaagacatg
     4801 agtacaggcc cttctttttc aactaaagac aatacgcttg aatactttgt gaaagcttct
     4861 aataaacacg gcttttcact tgatatttca ttaaatgtaa acggcgctgt gatttccggt
     4921 accatgattt cagcaaaaga atactttgat tacttaagcg aaacgtttga agaaggcagt
     4981 gaagtggctc aggcgctaag cgaacaattc tctttagcaa gcgaagcgag cgaatcaaac
     5041 ggagaagcag aagcccattt tattcatttg aaaaatacaa agatttactg tggagacagt
     5101 aaatctactc cttctaaagg caaaatcttt tggagaggga aaatagcaga agtagacggg
     5161 tttttcttag gaaagatttc tgatgcaaaa tcaacgagta aaaagagttc ataaaaaacg
     5221 gcggggtgat tgccccgccg ttttttagtg atgtgatgag atgtgcagct tcttttttcg
     5281 atacatgagc ggagtcatct tcatttcttt tcgaaactga ttaataaaat agcttgtact
     5341 attaaaccct acttgatagg cgacttccgt cacatttgct tctgcttgct gcagtccgga
     5401 ttcgcgaacg cgtctcgaga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     5461 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     5521 ggggtttttt gctgaaagga ggaactatat ccgactagta gcccgcctaa tgagcgggct
     5581 tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5641 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5701 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5761 cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac gagtgggata
     5821 catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg aagagcgttt
     5881 tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc gtgtagatgc
     5941 ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg ttgaatactc
     6001 cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat gtagtgccgc
     6061 cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa
     6121 ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg accgttggga
     6181 accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc ctgcggcaat
     6241 ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt cacggcaaca
     6301 attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt cggcacttcc
     6361 tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac ggggtatcat
     6421 tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca cgacgggtag
     6481 tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt cactgattaa
     6541 gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt ggacggaccg
     6601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7201 agcctatgga aa
//
