LOCUS       Exported                7672 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 7672)
  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..7672
                     /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..1435
                     /label=Bm1seq
     primer_bind     complement(1420..1435)
                     /label=gvpB_rev
     misc_feature    1435
                     /label=BR
     primer_bind     1436..1456
                     /label=gvpN_fwd
     misc_feature    1436
                     /label=RN
     misc_feature    1437..1445
                     /label=RBS
     CDS             1452..2378
                     /codon_start=1
                     /label=GvpN
                     /translation="MTVLTDKRKKGSGAFIQDDETKEVLSRALSYLKSGYSIHFTGPAG
                     GGKTSLARALAKKRKRPVMLMHGNHELNNKDLIGDFTGYTSKKVIDQYVRSVYKKDEQV
                     SENWQDGRLLEAVKNGYTLIYDEFTRSKPATNNIFLSILEEGVLPLYGVKMTDPFVRVH
                     PDFRVIFTSNPAEYAGVYDTQDALLDRLITMFIDYKDIDRETAILTEKTDVEEDEARTI
                     VTLVANVRNRSGDENSSGLSLRASLMIATLATQQDIPIDGSDEDFQTLCIDILHHPLTK
                     CLDEENAKSKAEKIILEECKNIDTEEK"
     primer_bind     1482..1507
                     /label=Bm1.1seqF
     misc_feature    1514
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     1884..1903
                     /label=Bm2seqF
     primer_bind     complement(2349..2378)
                     /label=gvpN_rev
     misc_feature    2378^2379
                     /label=NF
     primer_bind     2379..2398
                     /label=gvpF_fwd
     misc_feature    2379..2387
                     /label=RBS
     CDS             2393..3160
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLKVEEK"
     primer_bind     2626..2653
                     /label=Bm3seqF
     primer_bind     complement(3125..3145)
                     /label=gvpF_rev
     misc_feature    3145^3146
                     /label=FG
     primer_bind     3146..3168
                     /label=gvpG_fwd
     misc_feature    3150..3158
                     /label=RBS
     CDS             3162..3428
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKRNEES"
     primer_bind     3374..3401
                     /label=Bm4seqF
     primer_bind     complement(3392..3413)
                     /label=gvpG_rev
     misc_feature    3413^3414
                     /label=GL
     primer_bind     3414..3433
                     /label=gvpL_fwd
     misc_feature    3415..3423
                     /label=RBS
     CDS             3429..4238
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    3499
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3568
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4018
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4149..4175
                     /label=Bm5seqF
     primer_bind     complement(4222..4239)
                     /label=gvpL_rev
     misc_feature    4239^4240
                     /label=LS
     primer_bind     4240..4264
                     /label=gvpS_fwd
     misc_feature    4240..4248
                     /label=RBS
     CDS             4256..4543
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(4559..4578)
                     /label=gvpS_rev
     misc_feature    4578^4579
                     /label=SK
     primer_bind     4579..4594
                     /label=gvpK_fwd
     misc_feature    4600..4608
                     /label=RBS
     CDS             4615..4899
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4890..4905)
                     /label=gvpK_rev
     misc_feature    4905^4906
                     /label=KJ
     primer_bind     4906..4924
                     /label=gvpJ_fwd
     misc_feature    4909..4917
                     /label=RBS
     CDS             4921..5223
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    5046
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     5052..5073
                     /label=Bm6seqF
     primer_bind     complement(5218..5239)
                     /label=gvpJ_rev
     misc_feature    5239
                     /label=JT
     primer_bind     5240..5260
                     /label=gvpU_fwd
     misc_feature    5240
                     /label=TU
     misc_feature    5242..5250
                     /label=RBS
     CDS             5258..5674
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5597..5621
                     /label=GvpUseqF
     misc_feature    5827^5828
                     /label=UV
     primer_bind     5828..5845
                     /label=vector_fwd
     terminator      5944..5991
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6058..6149
                     /gene="bla"
                     /label=AmpR promoter
     RBS             6139..6144
                     /label=Emopec_63.3%
     CDS             6150..7010
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7084..7672
                     /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 gctttcagga
     1441 ggaacgtaaa aatgaccgtc ttaacagaca aaaggaaaaa aggcagtgga gcttttatac
     1501 aagatgacga gacaaaagag gttctttcaa gagcgctgag ctatttaaaa tccggctatt
     1561 ccattcattt tacaggtcct gccggcggag gcaaaacctc tttagcgcga gcgcttgcta
     1621 aaaagagaaa gcgtcctgta atgctgatgc acgggaatca cgagctcaac aacaaagatt
     1681 taattggcga ttttacggga tacacgagca aaaaagtaat cgaccagtac gttcgttctg
     1741 tctataaaaa agatgaacag gtgagtgaaa actggcagga tggccgattg cttgaagctg
     1801 taaaaaatgg ctatacgctg atttacgacg aatttactcg ttctaagcct gcgacgaata
     1861 atatctttct atcgatatta gaagaaggcg tgctgccgct gtatggagta aaaatgaccg
     1921 atccttttgt gcgcgtgcat cccgatttcc gcgtcatctt cacaagcaat ccagctgagt
     1981 atgccggcgt atatgatacg caagatgcgc ttctcgacag gttaattacc atgtttattg
     2041 attataaaga catcgacaga gagacagcga ttttaacgga gaaaacggac gtagaagaag
     2101 atgaagcgcg cacaattgta acgctcgtag caaacgtgcg aaaccgctct ggagacgaaa
     2161 acagcagcgg acttagcctg cgggcttcgc ttatgatcgc tacccttgcc acgcagcaag
     2221 acattcctat cgatggaagt gacgaagatt ttcaaacgtt atgtatcgat attttgcatc
     2281 atccgcttac caaatgtttg gatgaagaaa atgcaaaaag caaagccgaa aaaatcattt
     2341 tagaagaatg taagaatata gacactgaag aaaagtaaag gagcttgaaa acatgagtga
     2401 aacaaacgaa acaggtattt atatttttag cgccattcaa acggataaag acgaagaatt
     2461 tggcgccgtg gaagtagaag gaacaaaagc tgaaacattt ttgattcgct acaaagacgc
     2521 ggctatggta gcagctgaag taccgatgaa aatttatcat cctaatcgcc aaaatttatt
     2581 aatgcatcaa aacgcagtag cagcgattat ggacaagaac gatacggtta ttccaatcag
     2641 ctttgggaat gtattcaaat caaaagaaga cgtaaaagtt cttttggaaa acctttatcc
     2701 gcagtttgaa aagctgtttc cagcgatcaa aggaaaaatt gaagtcggtt taaaagtaat
     2761 tgggaaaaaa gaatggcttg agaaaaaagt aaacgaaaat cctgaacttg agaaagtatc
     2821 agcatccgta aaaggaaaat cagaagcagc cggttattat gagcgtattc aacttggagg
     2881 aatggctcaa aagatgttta cttccctgca aaaagaagtc aagacagatg tgttttctcc
     2941 gcttgaagaa gcagcggaag cagcaaaagc aaatgagcca acgggcgaaa cgatgctttt
     3001 aaacgcgtct ttcttaatta accgagaaga tgaagcgaag tttgatgaaa aagtaaatga
     3061 agcgcatgaa aactggaaag acaaagccga ttttcattac agcggtcctt ggcctgctta
     3121 taattttgtg aacattcgcc taaaagtaga agagaaataa cgtgcttcac aaattagtaa
     3181 ccgcacccat taaccttgta gtgaaaatcg gcgaaaaagt acaggaagaa gctgataaac
     3241 agctatatga ccttccgacg attcagcaaa agctcattca gcttcaaatg atgtttgagc
     3301 ttggtgaaat tccagaagaa gcgtttcaag aaaaagaaga tgaattgtta atgaggtacg
     3361 aaattgcgaa acgcagagaa attgaacaat gggaagagct aacacaaaaa agaaatgagg
     3421 aatcctagat gggagaatta ctgtatttat acggtttaat tccaacaaaa gaagcagcag
     3481 ccatagagcc gtttccatct tataaggggt ttgacggaga acattcactg tacccaattg
     3541 cgtttgatca ggtgacggct gtagtttcta agctggatgc tgacacctat tcagaaaaag
     3601 tgattcaaga aaaaatggag caggatatga gctggctgca ggaaaaagca tttcatcatc
     3661 acgaaacggt agccgctttg tacgaagaat ttacgatcat tccattaaaa ttttgcacca
     3721 tttataaagg tgaagaaagt ctgcaggcag ctattgagat taacaaagaa aagatagaga
     3781 attcactgac gctgcttcaa ggaaatgaag agtggaatgt gaaaatttac tgtgatgata
     3841 cagagcttaa aaaaggaatc agcgaaacga atgaaagcgt gaaagcgaaa aaacaagaaa
     3901 ttagtcactt atcaccagga agacagtttt ttgaaaagaa aaaaatagat cagctgattg
     3961 aaaaagaatt agagcttcac aaaaacaaag tgtgtgaaga gatacatgac aagctaaaag
     4021 aattatcgct ttatgactct gttaaaaaga attggagcaa agacgtaact ggcgcagctg
     4081 aacagatggc gtggaacagc gtgtttcttc tcccgtctct gcagattact aagttcgtaa
     4141 acgaaataga agagcttcag caaaggcttg aaaataaagg ctggaagttt gaagtgacgg
     4201 gaccatggcc gccctatcat ttctcgagct ttgcgtaaag tgaggaatta acattatgtc
     4261 tcttaaacaa tccatggaga ataaagatat tgctcttatt gatattttag atgtcatttt
     4321 agataaagga gtcgccatta aaggagactt aatcatttcc atagctggcg tcgatttagt
     4381 gtatttggat ttgcgggtgc ttatttcttc ggttgaaacg cttgtgcaag caaaagaagg
     4441 aaatcacaaa ccaatcactt ctgaacaatt tgataaacaa aaggaagagt taatggacgc
     4501 gaccggccag ccgagcaaat ggaccaatcc gcttggctcc tgacccttgc gagcagtcgt
     4561 atgctttctc gaattccgag cggttaagcg tgacagtcca ggaggaatta atggatgcaa
     4621 ccggtcagcc aagcaaatgg acgaatccac ttggatcctg atcaagctga acaaggctta
     4681 gcgcagcttg tgatgacagt tattgagcta ttgaggcaaa tcgttgaacg tcatgccatg
     4741 aggcgggtgg agggtggaac gttgacggac gaacaaattg aaaacttagg aattgcacta
     4801 atgaacttag aagaaaaaat ggacgagttg aaagaggtgt tcggtctgga tgcagaagat
     4861 ttaaatattg atcttggacc gctaggcagc ctgctttaag cggtcagtag gaggaacagt
     4921 atggcagtcg aacataatat gcagtcaagt acgattgtag atgtgctcga aaagattttg
     4981 gataaaggag tcgttatagc gggggacatc accgtaggaa ttgcagatgt cgagctatta
     5041 acgataaaga tccgcttgat tgtggcttcg gttgataagg caaaagaaat cggcatggac
     5101 tggtgggaaa atgatccgta tctcagttca aaaggagcca ataacaaagc gctcgaagaa
     5161 gaaaataaaa tgctgcatga gcggttaaaa acgcttgaag aaaaaataga aacgaaacgt
     5221 taaaaactgt acgctactta aaggaggaaa gaaagacatg agtacaggcc cttctttttc
     5281 aactaaagac aatacgcttg aatactttgt gaaagcttct aataaacacg gcttttcact
     5341 tgatatttca ttaaatgtaa acggcgctgt gatttccggt accatgattt cagcaaaaga
     5401 atactttgat tacttaagcg aaacgtttga agaaggcagt gaagtggctc aggcgctaag
     5461 cgaacaattc tctttagcaa gcgaagcgag cgaatcaaac ggagaagcag aagcccattt
     5521 tattcatttg aaaaatacaa agatttactg tggagacagt aaatctactc cttctaaagg
     5581 caaaatcttt tggagaggga aaatagcaga agtagacggg tttttcttag gaaagatttc
     5641 tgatgcaaaa tcaacgagta aaaagagttc ataaaaaacg gcggggtgat tgccccgccg
     5701 ttttttagtg atgtgatgag atgtgcagct tcttttttcg atacatgagc ggagtcatct
     5761 tcatttcttt tcgaaactga ttaataaaat agcttgtact attaaaccct acttgatagg
     5821 cgacttccgt cacatttgct tctgcttgct gcagtccgga ttcgcgaacg cgtctcgaga
     5881 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     5941 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     6001 ggaactatat ccgactagta gcccgcctaa tgagcgggct tttttttaat tcccctattt
     6061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag
     6241 taaaagacgc agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata
     6301 gcggtaaaat ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca
     6361 aagttctgct atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac
     6421 gacgaataca ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc
     6481 ttacggacgg aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca
     6541 ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc
     6601 acaatatggg agaccatgta actcgccttg accgttggga accagaactg aatgaagcca
     6661 taccaaacga cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac
     6721 tattaactgg cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag
     6781 cggataaagt tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg
     6841 ataaatctgg ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg
     6901 gtaagccttc ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac
     6961 gaaatagaca gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa
     7021 ttaaaggctc cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc
     7081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7201 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aa
//
