LOCUS       Exported                7725 bp ds-DNA     circular SYN 27-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7725)
  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..7725
                     /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    2685^2686
                     /label=NF
     primer_bind     2686..2705
                     /label=gvpF_fwd
     misc_feature    2686..2694
                     /label=RBS
     CDS             2700..3452
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLK"
     primer_bind     2933..2960
                     /label=Bm3seqF
     primer_bind     complement(3432..3452)
                     /label=gvpF_rev
     primer_bind     complement(3435..3465)
                     /label=FL.fix_rev
     misc_feature    3456..3460
                     /label=RBS
     CDS             3456..3460
                     /codon_start=1
                     /label=GvpG
                     /translation="E"
     primer_bind     3466..3495
                     /label=FL.fix_fwd
     CDS             3475..3481
                     /codon_start=1
                     /label=GvpG
                     /translation="S"
     misc_feature    3475..3476
                     /label=RBS
     CDS             3482..4291
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    3552
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3621
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4071
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4202..4228
                     /label=Bm5seqF
     primer_bind     complement(4275..4292)
                     /label=gvpL_rev
     misc_feature    4292^4293
                     /label=LS
     primer_bind     4293..4317
                     /label=gvpS_fwd
     misc_feature    4293..4301
                     /label=RBS
     CDS             4309..4596
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(4612..4631)
                     /label=gvpS_rev
     misc_feature    4631^4632
                     /label=SK
     primer_bind     4632..4647
                     /label=gvpK_fwd
     misc_feature    4653..4661
                     /label=RBS
     CDS             4668..4952
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4943..4958)
                     /label=gvpK_rev
     misc_feature    4958^4959
                     /label=KJ
     primer_bind     4959..4977
                     /label=gvpJ_fwd
     misc_feature    4962..4970
                     /label=RBS
     CDS             4974..5276
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    5099
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     5105..5126
                     /label=Bm6seqF
     primer_bind     complement(5271..5292)
                     /label=gvpJ_rev
     misc_feature    5292
                     /label=JT
     primer_bind     5293..5313
                     /label=gvpU_fwd
     misc_feature    5293
                     /label=TU
     misc_feature    5295..5303
                     /label=RBS
     CDS             5311..5727
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5650..5674
                     /label=GvpUseqF
     misc_feature    5880^5881
                     /label=UV
     primer_bind     5881..5898
                     /label=vector_fwd
     terminator      5997..6044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6111..6202
                     /gene="bla"
                     /label=AmpR promoter
     RBS             6192..6197
                     /label=Emopec_63.3%
     CDS             6203..7063
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7137..7725
                     /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 agtaaaggag cttgaaaaca
     2701 tgagtgaaac aaacgaaaca ggtatttata tttttagcgc cattcaaacg gataaagacg
     2761 aagaatttgg cgccgtggaa gtagaaggaa caaaagctga aacatttttg attcgctaca
     2821 aagacgcggc tatggtagca gctgaagtac cgatgaaaat ttatcatcct aatcgccaaa
     2881 atttattaat gcatcaaaac gcagtagcag cgattatgga caagaacgat acggttattc
     2941 caatcagctt tgggaatgta ttcaaatcaa aagaagacgt aaaagttctt ttggaaaacc
     3001 tttatccgca gtttgaaaag ctgtttccag cgatcaaagg aaaaattgaa gtcggtttaa
     3061 aagtaattgg gaaaaaagaa tggcttgaga aaaaagtaaa cgaaaatcct gaacttgaga
     3121 aagtatcagc atccgtaaaa ggaaaatcag aagcagccgg ttattatgag cgtattcaac
     3181 ttggaggaat ggctcaaaag atgtttactt ccctgcaaaa agaagtcaag acagatgtgt
     3241 tttctccgct tgaagaagca gcggaagcag caaaagcaaa tgagccaacg ggcgaaacga
     3301 tgcttttaaa cgcgtctttc ttaattaacc gagaagatga agcgaagttt gatgaaaaag
     3361 taaatgaagc gcatgaaaac tggaaagaca aagccgattt tcattacagc ggtccttggc
     3421 ctgcttataa ttttgtgaac attcgcctaa aagtagagga aaaataatca ggagatccta
     3481 gatgggagaa ttactgtatt tatacggttt aattccaaca aaagaagcag cagccataga
     3541 gccgtttcca tcttataagg ggtttgacgg agaacattca ctgtacccaa ttgcgtttga
     3601 tcaggtgacg gctgtagttt ctaagctgga tgctgacacc tattcagaaa aagtgattca
     3661 agaaaaaatg gagcaggata tgagctggct gcaggaaaaa gcatttcatc atcacgaaac
     3721 ggtagccgct ttgtacgaag aatttacgat cattccatta aaattttgca ccatttataa
     3781 aggtgaagaa agtctgcagg cagctattga gattaacaaa gaaaagatag agaattcact
     3841 gacgctgctt caaggaaatg aagagtggaa tgtgaaaatt tactgtgatg atacagagct
     3901 taaaaaagga atcagcgaaa cgaatgaaag cgtgaaagcg aaaaaacaag aaattagtca
     3961 cttatcacca ggaagacagt tttttgaaaa gaaaaaaata gatcagctga ttgaaaaaga
     4021 attagagctt cacaaaaaca aagtgtgtga agagatacat gacaagctaa aagaattatc
     4081 gctttatgac tctgttaaaa agaattggag caaagacgta actggcgcag ctgaacagat
     4141 ggcgtggaac agcgtgtttc ttctcccgtc tctgcagatt actaagttcg taaacgaaat
     4201 agaagagctt cagcaaaggc ttgaaaataa aggctggaag tttgaagtga cgggaccatg
     4261 gccgccctat catttctcga gctttgcgta aagtgaggaa ttaacattat gtctcttaaa
     4321 caatccatgg agaataaaga tattgctctt attgatattt tagatgtcat tttagataaa
     4381 ggagtcgcca ttaaaggaga cttaatcatt tccatagctg gcgtcgattt agtgtatttg
     4441 gatttgcggg tgcttatttc ttcggttgaa acgcttgtgc aagcaaaaga aggaaatcac
     4501 aaaccaatca cttctgaaca atttgataaa caaaaggaag agttaatgga cgcgaccggc
     4561 cagccgagca aatggaccaa tccgcttggc tcctgaccct tgcgagcagt cgtatgcttt
     4621 ctcgaattcc gagcggttaa gcgtgacagt ccaggaggaa ttaatggatg caaccggtca
     4681 gccaagcaaa tggacgaatc cacttggatc ctgatcaagc tgaacaaggc ttagcgcagc
     4741 ttgtgatgac agttattgag ctattgaggc aaatcgttga acgtcatgcc atgaggcggg
     4801 tggagggtgg aacgttgacg gacgaacaaa ttgaaaactt aggaattgca ctaatgaact
     4861 tagaagaaaa aatggacgag ttgaaagagg tgttcggtct ggatgcagaa gatttaaata
     4921 ttgatcttgg accgctaggc agcctgcttt aagcggtcag taggaggaac agtatggcag
     4981 tcgaacataa tatgcagtca agtacgattg tagatgtgct cgaaaagatt ttggataaag
     5041 gagtcgttat agcgggggac atcaccgtag gaattgcaga tgtcgagcta ttaacgataa
     5101 agatccgctt gattgtggct tcggttgata aggcaaaaga aatcggcatg gactggtggg
     5161 aaaatgatcc gtatctcagt tcaaaaggag ccaataacaa agcgctcgaa gaagaaaata
     5221 aaatgctgca tgagcggtta aaaacgcttg aagaaaaaat agaaacgaaa cgttaaaaac
     5281 tgtacgctac ttaaaggagg aaagaaagac atgagtacag gcccttcttt ttcaactaaa
     5341 gacaatacgc ttgaatactt tgtgaaagct tctaataaac acggcttttc acttgatatt
     5401 tcattaaatg taaacggcgc tgtgatttcc ggtaccatga tttcagcaaa agaatacttt
     5461 gattacttaa gcgaaacgtt tgaagaaggc agtgaagtgg ctcaggcgct aagcgaacaa
     5521 ttctctttag caagcgaagc gagcgaatca aacggagaag cagaagccca ttttattcat
     5581 ttgaaaaata caaagattta ctgtggagac agtaaatcta ctccttctaa aggcaaaatc
     5641 ttttggagag ggaaaatagc agaagtagac gggtttttct taggaaagat ttctgatgca
     5701 aaatcaacga gtaaaaagag ttcataaaaa acggcggggt gattgccccg ccgtttttta
     5761 gtgatgtgat gagatgtgca gcttcttttt tcgatacatg agcggagtca tcttcatttc
     5821 ttttcgaaac tgattaataa aatagcttgt actattaaac cctacttgat aggcgacttc
     5881 cgtcacattt gcttctgctt gctgcagtcc ggattcgcga acgcgtctcg agagatccgg
     5941 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     6001 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     6061 tatccgacta gtagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt
     6121 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6181 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6241 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga
     6301 cgcagaggac caattggggg cacgagtggg atacatagaa ctggacttga atagcggtaa
     6361 aatccttgag agttttcgcc ctgaagagcg ttttccaatg atgagcactt tcaaagttct
     6421 gctatgtgga gcagtattat cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat
     6481 acactattcg cagaatgact tggttgaata ctccccagtg acagaaaagc accttacgga
     6541 cggaatgacg gtaagagaat tatgtagtgc cgccataacg atgagtgata acactgcggc
     6601 gaacttactt ctgacaacca tcggtggacc gaaggaatta accgcttttt tgcacaatat
     6661 gggagaccat gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa
     6721 cgacgagcga gacaccacaa tgcctgcggc aatggcaaca acattacgca aactattaac
     6781 tggcgaacta cttactctgg cttcacggca acaattaata gactggcttg aagcggataa
     6841 agttgcagga ccactactgc gttcggcact tcctgctggc tggtttattg ctgataaatc
     6901 tggggcagga gagcgtggtt cacggggtat cattgccgca cttggaccag atggtaagcc
     6961 ttcccgtatc gtagttatct acacgacggg tagtcaggca actatggacg aacgaaatag
     7021 acagattgct gaaatagggg cttcactgat taagcattgg taaaccgata caattaaagg
     7081 ctccttttgg agcctttttt tttggacgga ccggtagaaa agatcaaagg atcttcttga
     7141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7261 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaa
//
