LOCUS       Exported                7762 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 7762)
  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..7762
                     /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..3413
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKR"
     primer_bind     3374..3401
                     /label=Bm4seqF
     primer_bind     complement(3392..3424)
                     /label=GS.fix_rev
     primer_bind     complement(3392..3413)
                     /label=gvpG_rev
     primer_bind     3425..3459
                     /label=GS.fix_fwd
     primer_bind     3436..3450
                     /label=gvpS_fwd
     CDS             3442..3729
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(3745..3764)
                     /label=gvpS_rev
     misc_feature    3764^3765
                     /label=SK
     primer_bind     3765..3780
                     /label=gvpK_fwd
     misc_feature    3786..3794
                     /label=RBS
     CDS             3801..4085
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4076..4091)
                     /label=gvpK_rev
     misc_feature    4091^4092
                     /label=KJ
     primer_bind     4092..4110
                     /label=gvpJ_fwd
     misc_feature    4095..4103
                     /label=RBS
     CDS             4107..4409
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    4232
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4238..4259
                     /label=Bm6seqF
     primer_bind     complement(4404..4425)
                     /label=gvpJ_rev
     misc_feature    4425^4426
                     /label=JT
     primer_bind     4426..4449
                     /label=gvpT_fwd
     misc_feature    4432..4440
                     /label=RBS
     CDS             4446..5324
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4467..4493
                     /label=Bm6.1seq
     misc_feature    4770
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4833
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4907..4933
                     /label=Bm7seqF
     primer_bind     complement(5312..5329)
                     /label=gvpT_rev
     misc_feature    5329^5330
                     /label=TU
     primer_bind     5330..5350
                     /label=gvpU_fwd
     misc_feature    5332..5340
                     /label=RBS
     CDS             5348..5764
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5687..5711
                     /label=GvpUseqF
     misc_feature    5917^5918
                     /label=UV
     primer_bind     5918..5935
                     /label=vector_fwd
     terminator      6034..6081
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6148..6239
                     /gene="bla"
                     /label=AmpR promoter
     RBS             6229..6234
                     /label=Emopec_63.3%
     CDS             6240..7100
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7174..7762
                     /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 agaaatgaag
     3421 aatcctaata ggagaaacat tatgtctctt aaacaatcca tggagaataa agatattgct
     3481 cttattgata ttttagatgt cattttagat aaaggagtcg ccattaaagg agacttaatc
     3541 atttccatag ctggcgtcga tttagtgtat ttggatttgc gggtgcttat ttcttcggtt
     3601 gaaacgcttg tgcaagcaaa agaaggaaat cacaaaccaa tcacttctga acaatttgat
     3661 aaacaaaagg aagagttaat ggacgcgacc ggccagccga gcaaatggac caatccgctt
     3721 ggctcctgac ccttgcgagc agtcgtatgc tttctcgaat tccgagcggt taagcgtgac
     3781 agtccaggag gaattaatgg atgcaaccgg tcagccaagc aaatggacga atccacttgg
     3841 atcctgatca agctgaacaa ggcttagcgc agcttgtgat gacagttatt gagctattga
     3901 ggcaaatcgt tgaacgtcat gccatgaggc gggtggaggg tggaacgttg acggacgaac
     3961 aaattgaaaa cttaggaatt gcactaatga acttagaaga aaaaatggac gagttgaaag
     4021 aggtgttcgg tctggatgca gaagatttaa atattgatct tggaccgcta ggcagcctgc
     4081 tttaagcggt cagtaggagg aacagtatgg cagtcgaaca taatatgcag tcaagtacga
     4141 ttgtagatgt gctcgaaaag attttggata aaggagtcgt tatagcgggg gacatcaccg
     4201 taggaattgc agatgtcgag ctattaacga taaagatccg cttgattgtg gcttcggttg
     4261 ataaggcaaa agaaatcggc atggactggt gggaaaatga tccgtatctc agttcaaaag
     4321 gagccaataa caaagcgctc gaagaagaaa ataaaatgct gcatgagcgg ttaaaaacgc
     4381 ttgaagaaaa aatagaaacg aaacgttaaa aactgtacgc tacttaaaaa atggagggat
     4441 ttacaatggc aactgaaaca aaattagata acacacaggc agaaaacaag gaaaataaaa
     4501 atgcggaaaa cggttcaaaa gaaaagaacg gttcaaaagc aagcaaaaca acaagcagcg
     4561 ggccaatcaa acgagcggta gcaggaggca tcatcggtgc aacgattgga tatgtatcga
     4621 ctcctgaaaa tcgaaaaagt ctccttgacc gcattgatac agacgaatta aaaagcaaag
     4681 catctgattt aggaacaaag gtaaaagaaa aatcaaaaag cagcgtggcc agcctgaaaa
     4741 catctgcggg aagcttgttt aaaaaagata aagataaatc aaaagatgat gaagaaaacg
     4801 taaattcttc tagtagcgaa acagaagacg ataacgttca agagtacgac gagttaaaag
     4861 aagaaaatca aactcttcaa gatcgcttat cacagcttga agaaaaaatg aacatgcttg
     4921 ttgagcttag cctcaataaa aatcaagacg aagaagcgga agatacagat tccgacgaag
     4981 aagagaacga tgagaacgat gaaaacgatg aaaacgagca ggacgatgaa aacgaagaag
     5041 aaacatctaa gccacgtaaa aaggataaaa aagaagctga ggaagaagaa agtgaaagtg
     5101 acgaagacag cgaggaagaa gaggaagatt ctcgctcaaa caaaaaaaat aaaaaagtaa
     5161 aaacagaaga agaagacgaa gatgaaagcg aagaagaaaa aaaggaagcg aaaccaaaaa
     5221 agtcaacagc taaaaaatca aaaaatacaa aagcaaagaa aaacacggac gaagaagatg
     5281 atgaagcaac atctctttct agtgaagacg atacaacagc ctaagacgta aaggaggaaa
     5341 gaaagacatg agtacaggcc cttctttttc aactaaagac aatacgcttg aatactttgt
     5401 gaaagcttct aataaacacg gcttttcact tgatatttca ttaaatgtaa acggcgctgt
     5461 gatttccggt accatgattt cagcaaaaga atactttgat tacttaagcg aaacgtttga
     5521 agaaggcagt gaagtggctc aggcgctaag cgaacaattc tctttagcaa gcgaagcgag
     5581 cgaatcaaac ggagaagcag aagcccattt tattcatttg aaaaatacaa agatttactg
     5641 tggagacagt aaatctactc cttctaaagg caaaatcttt tggagaggga aaatagcaga
     5701 agtagacggg tttttcttag gaaagatttc tgatgcaaaa tcaacgagta aaaagagttc
     5761 ataaaaaacg gcggggtgat tgccccgccg ttttttagtg atgtgatgag atgtgcagct
     5821 tcttttttcg atacatgagc ggagtcatct tcatttcttt tcgaaactga ttaataaaat
     5881 agcttgtact attaaaccct acttgatagg cgacttccgt cacatttgct tctgcttgct
     5941 gcagtccgga ttcgcgaacg cgtctcgaga gatccggctg ctaacaaagc ccgaaaggaa
     6001 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     6061 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccgactagta gcccgcctaa
     6121 tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt
     6181 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6241 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6301 tttttgctca cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa ttgggggcac
     6361 gagtgggata catagaactg gacttgaata gcggtaaaat ccttgagagt tttcgccctg
     6421 aagagcgttt tccaatgatg agcactttca aagttctgct atgtggagca gtattatccc
     6481 gtgtagatgc ggggcaagag caactcggac gacgaataca ctattcgcag aatgacttgg
     6541 ttgaatactc cccagtgaca gaaaagcacc ttacggacgg aatgacggta agagaattat
     6601 gtagtgccgc cataacgatg agtgataaca ctgcggcgaa cttacttctg acaaccatcg
     6661 gtggaccgaa ggaattaacc gcttttttgc acaatatggg agaccatgta actcgccttg
     6721 accgttggga accagaactg aatgaagcca taccaaacga cgagcgagac accacaatgc
     6781 ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg cgaactactt actctggctt
     6841 cacggcaaca attaatagac tggcttgaag cggataaagt tgcaggacca ctactgcgtt
     6901 cggcacttcc tgctggctgg tttattgctg ataaatctgg ggcaggagag cgtggttcac
     6961 ggggtatcat tgccgcactt ggaccagatg gtaagccttc ccgtatcgta gttatctaca
     7021 cgacgggtag tcaggcaact atggacgaac gaaatagaca gattgctgaa ataggggctt
     7081 cactgattaa gcattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt
     7141 ggacggaccg gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7321 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7741 aggggggcgg agcctatgga aa
//
