LOCUS       Exported                7407 bp ds-DNA     circular SYN 15-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7407)
  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..7407
                     /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
                     /label=VB
     primer_bind     1101..1123
                     /label=gvpR_fwd
     primer_bind     1101
                     /label=Bm1seq
     misc_feature    1101
                     /label=BR
     misc_feature    1114..1122
                     /label=RBS
     CDS             1129..1395
                     /codon_start=1
                     /label=GvpR
                     /translation="MEIKKIMQAVNDFFGEHVAPPHKITSVEATEDEGWRVIVEVIEER
                     EYMKKYAKDEMLGTYECFVNKEKEVISFKRLDVRYRSAIGIEA"
     primer_bind     complement(1386..1407)
                     /label=gvpR_rev
     misc_feature    1407^1408
                     /label=RN
     primer_bind     1408..1428
                     /label=gvpN_fwd
     misc_feature    1409..1417
                     /label=RBS
     CDS             1424..2350
                     /codon_start=1
                     /label=GvpN
                     /translation="MTVLTDKRKKGSGAFIQDDETKEVLSRALSYLKSGYSIHFTGPAG
                     GGKTSLARALAKKRKRPVMLMHGNHELNNKDLIGDFTGYTSKKVIDQYVRSVYKKDEQV
                     SENWQDGRLLEAVKNGYTLIYDEFTRSKPATNNIFLSILEEGVLPLYGVKMTDPFVRVH
                     PDFRVIFTSNPAEYAGVYDTQDALLDRLITMFIDYKDIDRETAILTEKTDVEEDEARTI
                     VTLVANVRNRSGDENSSGLSLRASLMIATLATQQDIPIDGSDEDFQTLCIDILHHPLTK
                     CLDEENAKSKAEKIILEECKNIDTEEK"
     primer_bind     1454..1479
                     /label=Bm1.1seqF
     misc_feature    1486
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     1856..1875
                     /label=Bm2seqF
     primer_bind     complement(2321..2350)
                     /label=gvpN_rev
     misc_feature    2350^2351
                     /label=NF
     primer_bind     2351..2370
                     /label=gvpF_fwd
     misc_feature    2351..2359
                     /label=RBS
     CDS             2365..3132
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLKVEEK"
     primer_bind     2598..2625
                     /label=Bm3seqF
     primer_bind     complement(3097..3117)
                     /label=gvpF_rev
     misc_feature    3117^3118
                     /label=FG
     primer_bind     3118..3140
                     /label=gvpG_fwd
     misc_feature    3122..3130
                     /label=RBS
     CDS             3134..3385
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKR"
     primer_bind     3346..3373
                     /label=Bm4seqF
     primer_bind     complement(3364..3396)
                     /label=GS.fix_rev
     primer_bind     complement(3364..3385)
                     /label=gvpG_rev
     primer_bind     3397..3431
                     /label=GS.fix_fwd
     primer_bind     3408..3422
                     /label=gvpS_fwd
     CDS             3414..3701
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(3717..3736)
                     /label=gvpS_rev
     misc_feature    3736
                     /label=SK
     primer_bind     3737..3755
                     /label=gvpJ_fwd
     misc_feature    3737
                     /label=KJ
     misc_feature    3740..3748
                     /label=RBS
     CDS             3752..4054
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    3877
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3883..3904
                     /label=Bm6seqF
     primer_bind     complement(4049..4070)
                     /label=gvpJ_rev
     misc_feature    4070^4071
                     /label=JT
     primer_bind     4071..4094
                     /label=gvpT_fwd
     misc_feature    4077..4085
                     /label=RBS
     CDS             4091..4969
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4112..4138
                     /label=Bm6.1seq
     misc_feature    4415
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4478
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4552..4578
                     /label=Bm7seqF
     primer_bind     complement(4957..4974)
                     /label=gvpT_rev
     misc_feature    4974^4975
                     /label=TU
     primer_bind     4975..4995
                     /label=gvpU_fwd
     misc_feature    4977..4985
                     /label=RBS
     CDS             4993..5409
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5332..5356
                     /label=GvpUseqF
     misc_feature    5562^5563
                     /label=UV
     primer_bind     5563..5580
                     /label=vector_fwd
     terminator      5679..5726
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5793..5884
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5874..5879
                     /label=Emopec_63.3%
     CDS             5885..6745
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6819..7407
                     /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 tagatctaac tattggaggc tactaaaaat ggaaattaaa
     1141 aaaattatgc aagccgtgaa cgactttttc ggtgaacacg tagctcctcc tcataaaatt
     1201 acctcggtgg aagctactga agatgaaggt tggagagtta ttgttgaagt cattgaagaa
     1261 cgagaatata tgaaaaaata cgccaaagat gaaatgctcg gaacgtacga gtgctttgta
     1321 aataaagaaa aagaagtcat ttcattcaaa cgactcgacg tcagatatag aagcgccatt
     1381 ggcattgaag cataaacgat aagatggcag gaggaacgta aaaatgaccg tcttaacaga
     1441 caaaaggaaa aaaggcagtg gagcttttat acaagatgac gagacaaaag aggttctttc
     1501 aagagcgctg agctatttaa aatccggcta ttccattcat tttacaggtc ctgccggcgg
     1561 aggcaaaacc tctttagcgc gagcgcttgc taaaaagaga aagcgtcctg taatgctgat
     1621 gcacgggaat cacgagctca acaacaaaga tttaattggc gattttacgg gatacacgag
     1681 caaaaaagta atcgaccagt acgttcgttc tgtctataaa aaagatgaac aggtgagtga
     1741 aaactggcag gatggccgat tgcttgaagc tgtaaaaaat ggctatacgc tgatttacga
     1801 cgaatttact cgttctaagc ctgcgacgaa taatatcttt ctatcgatat tagaagaagg
     1861 cgtgctgccg ctgtatggag taaaaatgac cgatcctttt gtgcgcgtgc atcccgattt
     1921 ccgcgtcatc ttcacaagca atccagctga gtatgccggc gtatatgata cgcaagatgc
     1981 gcttctcgac aggttaatta ccatgtttat tgattataaa gacatcgaca gagagacagc
     2041 gattttaacg gagaaaacgg acgtagaaga agatgaagcg cgcacaattg taacgctcgt
     2101 agcaaacgtg cgaaaccgct ctggagacga aaacagcagc ggacttagcc tgcgggcttc
     2161 gcttatgatc gctacccttg ccacgcagca agacattcct atcgatggaa gtgacgaaga
     2221 ttttcaaacg ttatgtatcg atattttgca tcatccgctt accaaatgtt tggatgaaga
     2281 aaatgcaaaa agcaaagccg aaaaaatcat tttagaagaa tgtaagaata tagacactga
     2341 agaaaagtaa aggagcttga aaacatgagt gaaacaaacg aaacaggtat ttatattttt
     2401 agcgccattc aaacggataa agacgaagaa tttggcgccg tggaagtaga aggaacaaaa
     2461 gctgaaacat ttttgattcg ctacaaagac gcggctatgg tagcagctga agtaccgatg
     2521 aaaatttatc atcctaatcg ccaaaattta ttaatgcatc aaaacgcagt agcagcgatt
     2581 atggacaaga acgatacggt tattccaatc agctttggga atgtattcaa atcaaaagaa
     2641 gacgtaaaag ttcttttgga aaacctttat ccgcagtttg aaaagctgtt tccagcgatc
     2701 aaaggaaaaa ttgaagtcgg tttaaaagta attgggaaaa aagaatggct tgagaaaaaa
     2761 gtaaacgaaa atcctgaact tgagaaagta tcagcatccg taaaaggaaa atcagaagca
     2821 gccggttatt atgagcgtat tcaacttgga ggaatggctc aaaagatgtt tacttccctg
     2881 caaaaagaag tcaagacaga tgtgttttct ccgcttgaag aagcagcgga agcagcaaaa
     2941 gcaaatgagc caacgggcga aacgatgctt ttaaacgcgt ctttcttaat taaccgagaa
     3001 gatgaagcga agtttgatga aaaagtaaat gaagcgcatg aaaactggaa agacaaagcc
     3061 gattttcatt acagcggtcc ttggcctgct tataattttg tgaacattcg cctaaaagta
     3121 gaagagaaat aacgtgcttc acaaattagt aaccgcaccc attaaccttg tagtgaaaat
     3181 cggcgaaaaa gtacaggaag aagctgataa acagctatat gaccttccga cgattcagca
     3241 aaagctcatt cagcttcaaa tgatgtttga gcttggtgaa attccagaag aagcgtttca
     3301 agaaaaagaa gatgaattgt taatgaggta cgaaattgcg aaacgcagag aaattgaaca
     3361 atgggaagag ctaacacaaa aaagaaatga agaatcctaa taggagaaac attatgtctc
     3421 ttaaacaatc catggagaat aaagatattg ctcttattga tattttagat gtcattttag
     3481 ataaaggagt cgccattaaa ggagacttaa tcatttccat agctggcgtc gatttagtgt
     3541 atttggattt gcgggtgctt atttcttcgg ttgaaacgct tgtgcaagca aaagaaggaa
     3601 atcacaaacc aatcacttct gaacaatttg ataaacaaaa ggaagagtta atggacgcga
     3661 ccggccagcc gagcaaatgg accaatccgc ttggctcctg acccttgcga gcagtcgtat
     3721 gctttctcga attccgagta ggaggaacag tatggcagtc gaacataata tgcagtcaag
     3781 tacgattgta gatgtgctcg aaaagatttt ggataaagga gtcgttatag cgggggacat
     3841 caccgtagga attgcagatg tcgagctatt aacgataaag atccgcttga ttgtggcttc
     3901 ggttgataag gcaaaagaaa tcggcatgga ctggtgggaa aatgatccgt atctcagttc
     3961 aaaaggagcc aataacaaag cgctcgaaga agaaaataaa atgctgcatg agcggttaaa
     4021 aacgcttgaa gaaaaaatag aaacgaaacg ttaaaaactg tacgctactt aaaaaatgga
     4081 gggatttaca atggcaactg aaacaaaatt agataacaca caggcagaaa acaaggaaaa
     4141 taaaaatgcg gaaaacggtt caaaagaaaa gaacggttca aaagcaagca aaacaacaag
     4201 cagcgggcca atcaaacgag cggtagcagg aggcatcatc ggtgcaacga ttggatatgt
     4261 atcgactcct gaaaatcgaa aaagtctcct tgaccgcatt gatacagacg aattaaaaag
     4321 caaagcatct gatttaggaa caaaggtaaa agaaaaatca aaaagcagcg tggccagcct
     4381 gaaaacatct gcgggaagct tgtttaaaaa agataaagat aaatcaaaag atgatgaaga
     4441 aaacgtaaat tcttctagta gcgaaacaga agacgataac gttcaagagt acgacgagtt
     4501 aaaagaagaa aatcaaactc ttcaagatcg cttatcacag cttgaagaaa aaatgaacat
     4561 gcttgttgag cttagcctca ataaaaatca agacgaagaa gcggaagata cagattccga
     4621 cgaagaagag aacgatgaga acgatgaaaa cgatgaaaac gagcaggacg atgaaaacga
     4681 agaagaaaca tctaagccac gtaaaaagga taaaaaagaa gctgaggaag aagaaagtga
     4741 aagtgacgaa gacagcgagg aagaagagga agattctcgc tcaaacaaaa aaaataaaaa
     4801 agtaaaaaca gaagaagaag acgaagatga aagcgaagaa gaaaaaaagg aagcgaaacc
     4861 aaaaaagtca acagctaaaa aatcaaaaaa tacaaaagca aagaaaaaca cggacgaaga
     4921 agatgatgaa gcaacatctc tttctagtga agacgataca acagcctaag acgtaaagga
     4981 ggaaagaaag acatgagtac aggcccttct ttttcaacta aagacaatac gcttgaatac
     5041 tttgtgaaag cttctaataa acacggcttt tcacttgata tttcattaaa tgtaaacggc
     5101 gctgtgattt ccggtaccat gatttcagca aaagaatact ttgattactt aagcgaaacg
     5161 tttgaagaag gcagtgaagt ggctcaggcg ctaagcgaac aattctcttt agcaagcgaa
     5221 gcgagcgaat caaacggaga agcagaagcc cattttattc atttgaaaaa tacaaagatt
     5281 tactgtggag acagtaaatc tactccttct aaaggcaaaa tcttttggag agggaaaata
     5341 gcagaagtag acgggttttt cttaggaaag atttctgatg caaaatcaac gagtaaaaag
     5401 agttcataaa aaacggcggg gtgattgccc cgccgttttt tagtgatgtg atgagatgtg
     5461 cagcttcttt tttcgataca tgagcggagt catcttcatt tcttttcgaa actgattaat
     5521 aaaatagctt gtactattaa accctacttg ataggcgact tccgtcacat ttgcttctgc
     5581 ttgctgcagt ccggattcgc gaacgcgtct cgagagatcc ggctgctaac aaagcccgaa
     5641 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5701 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgac tagtagcccg
     5761 cctaatgagc gggctttttt ttaattcccc tatttgttta tttttctaaa tacattcaaa
     5821 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5881 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5941 tcctgttttt gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg accaattggg
     6001 ggcacgagtg ggatacatag aactggactt gaatagcggt aaaatccttg agagttttcg
     6061 ccctgaagag cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt
     6121 atcccgtgta gatgcggggc aagagcaact cggacgacga atacactatt cgcagaatga
     6181 cttggttgaa tactccccag tgacagaaaa gcaccttacg gacggaatga cggtaagaga
     6241 attatgtagt gccgccataa cgatgagtga taacactgcg gcgaacttac ttctgacaac
     6301 catcggtgga ccgaaggaat taaccgcttt tttgcacaat atgggagacc atgtaactcg
     6361 ccttgaccgt tgggaaccag aactgaatga agccatacca aacgacgagc gagacaccac
     6421 aatgcctgcg gcaatggcaa caacattacg caaactatta actggcgaac tacttactct
     6481 ggcttcacgg caacaattaa tagactggct tgaagcggat aaagttgcag gaccactact
     6541 gcgttcggca cttcctgctg gctggtttat tgctgataaa tctggggcag gagagcgtgg
     6601 ttcacggggt atcattgccg cacttggacc agatggtaag ccttcccgta tcgtagttat
     6661 ctacacgacg ggtagtcagg caactatgga cgaacgaaat agacagattg ctgaaatagg
     6721 ggcttcactg attaagcatt ggtaaaccga tacaattaaa ggctcctttt ggagcctttt
     6781 tttttggacg gaccggtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6961 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7381 tcgtcagggg ggcggagcct atggaaa
//
