LOCUS       Exported                7302 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 7302)
  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..7302
                     /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..2953
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKR"
     primer_bind     2914..2941
                     /label=Bm4seqF
     primer_bind     complement(2932..2964)
                     /label=GS.fix_rev
     primer_bind     complement(2932..2953)
                     /label=gvpG_rev
     primer_bind     2965..2999
                     /label=GS.fix_fwd
     primer_bind     2976..2990
                     /label=gvpS_fwd
     CDS             2982..3269
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(3285..3304)
                     /label=gvpS_rev
     misc_feature    3304^3305
                     /label=SK
     primer_bind     3305..3320
                     /label=gvpK_fwd
     misc_feature    3326..3334
                     /label=RBS
     CDS             3341..3625
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(3616..3631)
                     /label=gvpK_rev
     misc_feature    3631^3632
                     /label=KJ
     primer_bind     3632..3650
                     /label=gvpJ_fwd
     misc_feature    3635..3643
                     /label=RBS
     CDS             3647..3949
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    3772
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3778..3799
                     /label=Bm6seqF
     primer_bind     complement(3944..3965)
                     /label=gvpJ_rev
     misc_feature    3965^3966
                     /label=JT
     primer_bind     3966..3989
                     /label=gvpT_fwd
     misc_feature    3972..3980
                     /label=RBS
     CDS             3986..4864
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4007..4033
                     /label=Bm6.1seq
     misc_feature    4310
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4373
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4447..4473
                     /label=Bm7seqF
     primer_bind     complement(4852..4869)
                     /label=gvpT_rev
     misc_feature    4869^4870
                     /label=TU
     primer_bind     4870..4890
                     /label=gvpU_fwd
     misc_feature    4872..4880
                     /label=RBS
     CDS             4888..5304
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5227..5251
                     /label=GvpUseqF
     misc_feature    5457^5458
                     /label=UV
     primer_bind     5458..5475
                     /label=vector_fwd
     terminator      5574..5621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5688..5779
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5769..5774
                     /label=Emopec_63.3%
     CDS             5780..6640
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6714..7302
                     /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 agaaatgaag aatcctaata ggagaaacat tatgtctctt aaacaatcca
     3001 tggagaataa agatattgct cttattgata ttttagatgt cattttagat aaaggagtcg
     3061 ccattaaagg agacttaatc atttccatag ctggcgtcga tttagtgtat ttggatttgc
     3121 gggtgcttat ttcttcggtt gaaacgcttg tgcaagcaaa agaaggaaat cacaaaccaa
     3181 tcacttctga acaatttgat aaacaaaagg aagagttaat ggacgcgacc ggccagccga
     3241 gcaaatggac caatccgctt ggctcctgac ccttgcgagc agtcgtatgc tttctcgaat
     3301 tccgagcggt taagcgtgac agtccaggag gaattaatgg atgcaaccgg tcagccaagc
     3361 aaatggacga atccacttgg atcctgatca agctgaacaa ggcttagcgc agcttgtgat
     3421 gacagttatt gagctattga ggcaaatcgt tgaacgtcat gccatgaggc gggtggaggg
     3481 tggaacgttg acggacgaac aaattgaaaa cttaggaatt gcactaatga acttagaaga
     3541 aaaaatggac gagttgaaag aggtgttcgg tctggatgca gaagatttaa atattgatct
     3601 tggaccgcta ggcagcctgc tttaagcggt cagtaggagg aacagtatgg cagtcgaaca
     3661 taatatgcag tcaagtacga ttgtagatgt gctcgaaaag attttggata aaggagtcgt
     3721 tatagcgggg gacatcaccg taggaattgc agatgtcgag ctattaacga taaagatccg
     3781 cttgattgtg gcttcggttg ataaggcaaa agaaatcggc atggactggt gggaaaatga
     3841 tccgtatctc agttcaaaag gagccaataa caaagcgctc gaagaagaaa ataaaatgct
     3901 gcatgagcgg ttaaaaacgc ttgaagaaaa aatagaaacg aaacgttaaa aactgtacgc
     3961 tacttaaaaa atggagggat ttacaatggc aactgaaaca aaattagata acacacaggc
     4021 agaaaacaag gaaaataaaa atgcggaaaa cggttcaaaa gaaaagaacg gttcaaaagc
     4081 aagcaaaaca acaagcagcg ggccaatcaa acgagcggta gcaggaggca tcatcggtgc
     4141 aacgattgga tatgtatcga ctcctgaaaa tcgaaaaagt ctccttgacc gcattgatac
     4201 agacgaatta aaaagcaaag catctgattt aggaacaaag gtaaaagaaa aatcaaaaag
     4261 cagcgtggcc agcctgaaaa catctgcggg aagcttgttt aaaaaagata aagataaatc
     4321 aaaagatgat gaagaaaacg taaattcttc tagtagcgaa acagaagacg ataacgttca
     4381 agagtacgac gagttaaaag aagaaaatca aactcttcaa gatcgcttat cacagcttga
     4441 agaaaaaatg aacatgcttg ttgagcttag cctcaataaa aatcaagacg aagaagcgga
     4501 agatacagat tccgacgaag aagagaacga tgagaacgat gaaaacgatg aaaacgagca
     4561 ggacgatgaa aacgaagaag aaacatctaa gccacgtaaa aaggataaaa aagaagctga
     4621 ggaagaagaa agtgaaagtg acgaagacag cgaggaagaa gaggaagatt ctcgctcaaa
     4681 caaaaaaaat aaaaaagtaa aaacagaaga agaagacgaa gatgaaagcg aagaagaaaa
     4741 aaaggaagcg aaaccaaaaa agtcaacagc taaaaaatca aaaaatacaa aagcaaagaa
     4801 aaacacggac gaagaagatg atgaagcaac atctctttct agtgaagacg atacaacagc
     4861 ctaagacgta aaggaggaaa gaaagacatg agtacaggcc cttctttttc aactaaagac
     4921 aatacgcttg aatactttgt gaaagcttct aataaacacg gcttttcact tgatatttca
     4981 ttaaatgtaa acggcgctgt gatttccggt accatgattt cagcaaaaga atactttgat
     5041 tacttaagcg aaacgtttga agaaggcagt gaagtggctc aggcgctaag cgaacaattc
     5101 tctttagcaa gcgaagcgag cgaatcaaac ggagaagcag aagcccattt tattcatttg
     5161 aaaaatacaa agatttactg tggagacagt aaatctactc cttctaaagg caaaatcttt
     5221 tggagaggga aaatagcaga agtagacggg tttttcttag gaaagatttc tgatgcaaaa
     5281 tcaacgagta aaaagagttc ataaaaaacg gcggggtgat tgccccgccg ttttttagtg
     5341 atgtgatgag atgtgcagct tcttttttcg atacatgagc ggagtcatct tcatttcttt
     5401 tcgaaactga ttaataaaat agcttgtact attaaaccct acttgatagg cgacttccgt
     5461 cacatttgct tctgcttgct gcagtccgga ttcgcgaacg cgtctcgaga gatccggctg
     5521 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     5581 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     5641 ccgactagta gcccgcctaa tgagcgggct tttttttaat tcccctattt gtttattttt
     5701 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5761 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5821 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag taaaagacgc
     5881 agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata gcggtaaaat
     5941 ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca aagttctgct
     6001 atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac gacgaataca
     6061 ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc ttacggacgg
     6121 aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca ctgcggcgaa
     6181 cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc acaatatggg
     6241 agaccatgta actcgccttg accgttggga accagaactg aatgaagcca taccaaacga
     6301 cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg
     6361 cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag cggataaagt
     6421 tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg ataaatctgg
     6481 ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg gtaagccttc
     6541 ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac gaaatagaca
     6601 gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa ttaaaggctc
     6661 cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc ttcttgagat
     6721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6841 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aa
//
