LOCUS       Exported                7686 bp ds-DNA     circular SYN 26-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  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..7686
                     /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
                     /label=RN
     primer_bind     1743..1762
                     /label=gvpF_fwd
     misc_feature    1743..1751
                     /label=RBS
     misc_feature    1743
                     /label=NF
     CDS             1757..2509
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLK"
     primer_bind     1990..2017
                     /label=Bm3seqF
     primer_bind     complement(2489..2509)
                     /label=gvpF_rev
     primer_bind     complement(2492..2522)
                     /label=FL.fix_rev
     misc_feature    2513..2517
                     /label=RBS
     CDS             2513..2517
                     /codon_start=1
                     /label=GvpG
                     /translation="E"
     primer_bind     2523..2552
                     /label=FL.fix_fwd
     CDS             2532..2538
                     /codon_start=1
                     /label=GvpG
                     /translation="S"
     misc_feature    2532..2533
                     /label=RBS
     CDS             2539..3348
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    2609
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    2678
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3128
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3259..3285
                     /label=Bm5seqF
     primer_bind     complement(3332..3349)
                     /label=gvpL_rev
     misc_feature    3349^3350
                     /label=LS
     primer_bind     3350..3374
                     /label=gvpS_fwd
     misc_feature    3350..3358
                     /label=RBS
     CDS             3366..3653
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(3669..3688)
                     /label=gvpS_rev
     misc_feature    3688^3689
                     /label=SK
     primer_bind     3689..3704
                     /label=gvpK_fwd
     misc_feature    3710..3718
                     /label=RBS
     CDS             3725..4009
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4000..4015)
                     /label=gvpK_rev
     misc_feature    4015^4016
                     /label=KJ
     primer_bind     4016..4034
                     /label=gvpJ_fwd
     misc_feature    4019..4027
                     /label=RBS
     CDS             4031..4333
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    4156
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4162..4183
                     /label=Bm6seqF
     primer_bind     complement(4328..4349)
                     /label=gvpJ_rev
     misc_feature    4349^4350
                     /label=JT
     primer_bind     4350..4373
                     /label=gvpT_fwd
     misc_feature    4356..4364
                     /label=RBS
     CDS             4370..5248
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4391..4417
                     /label=Bm6.1seq
     misc_feature    4694
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4757
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4831..4857
                     /label=Bm7seqF
     primer_bind     complement(5236..5253)
                     /label=gvpT_rev
     misc_feature    5253^5254
                     /label=TU
     primer_bind     5254..5274
                     /label=gvpU_fwd
     misc_feature    5256..5264
                     /label=RBS
     CDS             5272..5688
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5611..5635
                     /label=GvpUseqF
     misc_feature    5841^5842
                     /label=UV
     primer_bind     5842..5859
                     /label=vector_fwd
     terminator      5958..6005
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6072..6163
                     /gene="bla"
                     /label=AmpR promoter
     RBS             6153..6158
                     /label=Emopec_63.3%
     CDS             6164..7024
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7098..7686
                     /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 ggaggagctt gaaaacatga gtgaaacaaa cgaaacaggt atttatattt ttagcgccat
     1801 tcaaacggat aaagacgaag aatttggcgc cgtggaagta gaaggaacaa aagctgaaac
     1861 atttttgatt cgctacaaag acgcggctat ggtagcagct gaagtaccga tgaaaattta
     1921 tcatcctaat cgccaaaatt tattaatgca tcaaaacgca gtagcagcga ttatggacaa
     1981 gaacgatacg gttattccaa tcagctttgg gaatgtattc aaatcaaaag aagacgtaaa
     2041 agttcttttg gaaaaccttt atccgcagtt tgaaaagctg tttccagcga tcaaaggaaa
     2101 aattgaagtc ggtttaaaag taattgggaa aaaagaatgg cttgagaaaa aagtaaacga
     2161 aaatcctgaa cttgagaaag tatcagcatc cgtaaaagga aaatcagaag cagccggtta
     2221 ttatgagcgt attcaacttg gaggaatggc tcaaaagatg tttacttccc tgcaaaaaga
     2281 agtcaagaca gatgtgtttt ctccgcttga agaagcagcg gaagcagcaa aagcaaatga
     2341 gccaacgggc gaaacgatgc ttttaaacgc gtctttctta attaaccgag aagatgaagc
     2401 gaagtttgat gaaaaagtaa atgaagcgca tgaaaactgg aaagacaaag ccgattttca
     2461 ttacagcggt ccttggcctg cttataattt tgtgaacatt cgcctaaaag tagaggaaaa
     2521 ataatcagga gatcctagat gggagaatta ctgtatttat acggtttaat tccaacaaaa
     2581 gaagcagcag ccatagagcc gtttccatct tataaggggt ttgacggaga acattcactg
     2641 tacccaattg cgtttgatca ggtgacggct gtagtttcta agctggatgc tgacacctat
     2701 tcagaaaaag tgattcaaga aaaaatggag caggatatga gctggctgca ggaaaaagca
     2761 tttcatcatc acgaaacggt agccgctttg tacgaagaat ttacgatcat tccattaaaa
     2821 ttttgcacca tttataaagg tgaagaaagt ctgcaggcag ctattgagat taacaaagaa
     2881 aagatagaga attcactgac gctgcttcaa ggaaatgaag agtggaatgt gaaaatttac
     2941 tgtgatgata cagagcttaa aaaaggaatc agcgaaacga atgaaagcgt gaaagcgaaa
     3001 aaacaagaaa ttagtcactt atcaccagga agacagtttt ttgaaaagaa aaaaatagat
     3061 cagctgattg aaaaagaatt agagcttcac aaaaacaaag tgtgtgaaga gatacatgac
     3121 aagctaaaag aattatcgct ttatgactct gttaaaaaga attggagcaa agacgtaact
     3181 ggcgcagctg aacagatggc gtggaacagc gtgtttcttc tcccgtctct gcagattact
     3241 aagttcgtaa acgaaataga agagcttcag caaaggcttg aaaataaagg ctggaagttt
     3301 gaagtgacgg gaccatggcc gccctatcat ttctcgagct ttgcgtaaag tgaggaatta
     3361 acattatgtc tcttaaacaa tccatggaga ataaagatat tgctcttatt gatattttag
     3421 atgtcatttt agataaagga gtcgccatta aaggagactt aatcatttcc atagctggcg
     3481 tcgatttagt gtatttggat ttgcgggtgc ttatttcttc ggttgaaacg cttgtgcaag
     3541 caaaagaagg aaatcacaaa ccaatcactt ctgaacaatt tgataaacaa aaggaagagt
     3601 taatggacgc gaccggccag ccgagcaaat ggaccaatcc gcttggctcc tgacccttgc
     3661 gagcagtcgt atgctttctc gaattccgag cggttaagcg tgacagtcca ggaggaatta
     3721 atggatgcaa ccggtcagcc aagcaaatgg acgaatccac ttggatcctg atcaagctga
     3781 acaaggctta gcgcagcttg tgatgacagt tattgagcta ttgaggcaaa tcgttgaacg
     3841 tcatgccatg aggcgggtgg agggtggaac gttgacggac gaacaaattg aaaacttagg
     3901 aattgcacta atgaacttag aagaaaaaat ggacgagttg aaagaggtgt tcggtctgga
     3961 tgcagaagat ttaaatattg atcttggacc gctaggcagc ctgctttaag cggtcagtag
     4021 gaggaacagt atggcagtcg aacataatat gcagtcaagt acgattgtag atgtgctcga
     4081 aaagattttg gataaaggag tcgttatagc gggggacatc accgtaggaa ttgcagatgt
     4141 cgagctatta acgataaaga tccgcttgat tgtggcttcg gttgataagg caaaagaaat
     4201 cggcatggac tggtgggaaa atgatccgta tctcagttca aaaggagcca ataacaaagc
     4261 gctcgaagaa gaaaataaaa tgctgcatga gcggttaaaa acgcttgaag aaaaaataga
     4321 aacgaaacgt taaaaactgt acgctactta aaaaatggag ggatttacaa tggcaactga
     4381 aacaaaatta gataacacac aggcagaaaa caaggaaaat aaaaatgcgg aaaacggttc
     4441 aaaagaaaag aacggttcaa aagcaagcaa aacaacaagc agcgggccaa tcaaacgagc
     4501 ggtagcagga ggcatcatcg gtgcaacgat tggatatgta tcgactcctg aaaatcgaaa
     4561 aagtctcctt gaccgcattg atacagacga attaaaaagc aaagcatctg atttaggaac
     4621 aaaggtaaaa gaaaaatcaa aaagcagcgt ggccagcctg aaaacatctg cgggaagctt
     4681 gtttaaaaaa gataaagata aatcaaaaga tgatgaagaa aacgtaaatt cttctagtag
     4741 cgaaacagaa gacgataacg ttcaagagta cgacgagtta aaagaagaaa atcaaactct
     4801 tcaagatcgc ttatcacagc ttgaagaaaa aatgaacatg cttgttgagc ttagcctcaa
     4861 taaaaatcaa gacgaagaag cggaagatac agattccgac gaagaagaga acgatgagaa
     4921 cgatgaaaac gatgaaaacg agcaggacga tgaaaacgaa gaagaaacat ctaagccacg
     4981 taaaaaggat aaaaaagaag ctgaggaaga agaaagtgaa agtgacgaag acagcgagga
     5041 agaagaggaa gattctcgct caaacaaaaa aaataaaaaa gtaaaaacag aagaagaaga
     5101 cgaagatgaa agcgaagaag aaaaaaagga agcgaaacca aaaaagtcaa cagctaaaaa
     5161 atcaaaaaat acaaaagcaa agaaaaacac ggacgaagaa gatgatgaag caacatctct
     5221 ttctagtgaa gacgatacaa cagcctaaga cgtaaaggag gaaagaaaga catgagtaca
     5281 ggcccttctt tttcaactaa agacaatacg cttgaatact ttgtgaaagc ttctaataaa
     5341 cacggctttt cacttgatat ttcattaaat gtaaacggcg ctgtgatttc cggtaccatg
     5401 atttcagcaa aagaatactt tgattactta agcgaaacgt ttgaagaagg cagtgaagtg
     5461 gctcaggcgc taagcgaaca attctcttta gcaagcgaag cgagcgaatc aaacggagaa
     5521 gcagaagccc attttattca tttgaaaaat acaaagattt actgtggaga cagtaaatct
     5581 actccttcta aaggcaaaat cttttggaga gggaaaatag cagaagtaga cgggtttttc
     5641 ttaggaaaga tttctgatgc aaaatcaacg agtaaaaaga gttcataaaa aacggcgggg
     5701 tgattgcccc gccgtttttt agtgatgtga tgagatgtgc agcttctttt ttcgatacat
     5761 gagcggagtc atcttcattt cttttcgaaa ctgattaata aaatagcttg tactattaaa
     5821 ccctacttga taggcgactt ccgtcacatt tgcttctgct tgctgcagtc cggattcgcg
     5881 aacgcgtctc gagagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     5941 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     6001 ttttgctgaa aggaggaact atatccgact agtagcccgc ctaatgagcg ggcttttttt
     6061 taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6241 aacgctcgtg aaagtaaaag acgcagagga ccaattgggg gcacgagtgg gatacataga
     6301 actggacttg aatagcggta aaatccttga gagttttcgc cctgaagagc gttttccaat
     6361 gatgagcact ttcaaagttc tgctatgtgg agcagtatta tcccgtgtag atgcggggca
     6421 agagcaactc ggacgacgaa tacactattc gcagaatgac ttggttgaat actccccagt
     6481 gacagaaaag caccttacgg acggaatgac ggtaagagaa ttatgtagtg ccgccataac
     6541 gatgagtgat aacactgcgg cgaacttact tctgacaacc atcggtggac cgaaggaatt
     6601 aaccgctttt ttgcacaata tgggagacca tgtaactcgc cttgaccgtt gggaaccaga
     6661 actgaatgaa gccataccaa acgacgagcg agacaccaca atgcctgcgg caatggcaac
     6721 aacattacgc aaactattaa ctggcgaact acttactctg gcttcacggc aacaattaat
     6781 agactggctt gaagcggata aagttgcagg accactactg cgttcggcac ttcctgctgg
     6841 ctggtttatt gctgataaat ctggggcagg agagcgtggt tcacggggta tcattgccgc
     6901 acttggacca gatggtaagc cttcccgtat cgtagttatc tacacgacgg gtagtcaggc
     6961 aactatggac gaacgaaata gacagattgc tgaaataggg gcttcactga ttaagcattg
     7021 gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacgg accggtagaa
     7081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     7261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7681 tggaaa
//
