LOCUS       Exported                7352 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 7352)
  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..7352
                     /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..2524
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLKVEEK"
     primer_bind     1990..2017
                     /label=Bm3seqF
     primer_bind     complement(2489..2509)
                     /label=gvpF_rev
     misc_feature    2509^2510
                     /label=FG
     primer_bind     2510..2532
                     /label=gvpG_fwd
     misc_feature    2514..2522
                     /label=RBS
     CDS             2526..2792
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKRNEES"
     primer_bind     2738..2765
                     /label=Bm4seqF
     primer_bind     complement(2756..2777)
                     /label=gvpG_rev
     misc_feature    2777^2778
                     /label=GL
     primer_bind     2778..2797
                     /label=gvpL_fwd
     misc_feature    2779..2787
                     /label=RBS
     CDS             2793..3602
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    2863
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    2932
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    3382
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3513..3539
                     /label=Bm5seqF
     primer_bind     complement(3586..3603)
                     /label=gvpL_rev
     misc_feature    3603^3604
                     /label=LS
     primer_bind     3604..3628
                     /label=gvpS_fwd
     misc_feature    3604..3612
                     /label=RBS
     CDS             3620..3907
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(3923..3942)
                     /label=gvpS_rev
     misc_feature    3942^3943
                     /label=SK
     primer_bind     3943..3958
                     /label=gvpK_fwd
     misc_feature    3964..3972
                     /label=RBS
     CDS             3979..4263
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4254..4269)
                     /label=gvpK_rev
     misc_feature    4269^4270
                     /label=KJ
     primer_bind     4270..4288
                     /label=gvpJ_fwd
     misc_feature    4273..4281
                     /label=RBS
     CDS             4285..4587
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    4410
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4416..4437
                     /label=Bm6seqF
     primer_bind     complement(4582..4603)
                     /label=gvpJ_rev
     misc_feature    4603^4604
                     /label=JT
     primer_bind     4604..4627
                     /label=gvpT_fwd
     misc_feature    4610..4618
                     /label=RBS
     CDS             4624..5502
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4645..4671
                     /label=Bm6.1seq
     misc_feature    4948
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    5011
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     5085..5111
                     /label=Bm7seqF
     primer_bind     complement(5490..5507)
                     /label=gvpT_rev
     misc_feature    5507
                     /label=TU
     primer_bind     5508..5525
                     /label=vector_fwd
     misc_feature    5508
                     /label=UV
     terminator      5624..5671
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5738..5829
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5819..5824
                     /label=Emopec_63.3%
     CDS             5830..6690
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6764..7352
                     /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 tagaagagaa
     2521 ataacgtgct tcacaaatta gtaaccgcac ccattaacct tgtagtgaaa atcggcgaaa
     2581 aagtacagga agaagctgat aaacagctat atgaccttcc gacgattcag caaaagctca
     2641 ttcagcttca aatgatgttt gagcttggtg aaattccaga agaagcgttt caagaaaaag
     2701 aagatgaatt gttaatgagg tacgaaattg cgaaacgcag agaaattgaa caatgggaag
     2761 agctaacaca aaaaagaaat gaggaatcct agatgggaga attactgtat ttatacggtt
     2821 taattccaac aaaagaagca gcagccatag agccgtttcc atcttataag gggtttgacg
     2881 gagaacattc actgtaccca attgcgtttg atcaggtgac ggctgtagtt tctaagctgg
     2941 atgctgacac ctattcagaa aaagtgattc aagaaaaaat ggagcaggat atgagctggc
     3001 tgcaggaaaa agcatttcat catcacgaaa cggtagccgc tttgtacgaa gaatttacga
     3061 tcattccatt aaaattttgc accatttata aaggtgaaga aagtctgcag gcagctattg
     3121 agattaacaa agaaaagata gagaattcac tgacgctgct tcaaggaaat gaagagtgga
     3181 atgtgaaaat ttactgtgat gatacagagc ttaaaaaagg aatcagcgaa acgaatgaaa
     3241 gcgtgaaagc gaaaaaacaa gaaattagtc acttatcacc aggaagacag ttttttgaaa
     3301 agaaaaaaat agatcagctg attgaaaaag aattagagct tcacaaaaac aaagtgtgtg
     3361 aagagataca tgacaagcta aaagaattat cgctttatga ctctgttaaa aagaattgga
     3421 gcaaagacgt aactggcgca gctgaacaga tggcgtggaa cagcgtgttt cttctcccgt
     3481 ctctgcagat tactaagttc gtaaacgaaa tagaagagct tcagcaaagg cttgaaaata
     3541 aaggctggaa gtttgaagtg acgggaccat ggccgcccta tcatttctcg agctttgcgt
     3601 aaagtgagga attaacatta tgtctcttaa acaatccatg gagaataaag atattgctct
     3661 tattgatatt ttagatgtca ttttagataa aggagtcgcc attaaaggag acttaatcat
     3721 ttccatagct ggcgtcgatt tagtgtattt ggatttgcgg gtgcttattt cttcggttga
     3781 aacgcttgtg caagcaaaag aaggaaatca caaaccaatc acttctgaac aatttgataa
     3841 acaaaaggaa gagttaatgg acgcgaccgg ccagccgagc aaatggacca atccgcttgg
     3901 ctcctgaccc ttgcgagcag tcgtatgctt tctcgaattc cgagcggtta agcgtgacag
     3961 tccaggagga attaatggat gcaaccggtc agccaagcaa atggacgaat ccacttggat
     4021 cctgatcaag ctgaacaagg cttagcgcag cttgtgatga cagttattga gctattgagg
     4081 caaatcgttg aacgtcatgc catgaggcgg gtggagggtg gaacgttgac ggacgaacaa
     4141 attgaaaact taggaattgc actaatgaac ttagaagaaa aaatggacga gttgaaagag
     4201 gtgttcggtc tggatgcaga agatttaaat attgatcttg gaccgctagg cagcctgctt
     4261 taagcggtca gtaggaggaa cagtatggca gtcgaacata atatgcagtc aagtacgatt
     4321 gtagatgtgc tcgaaaagat tttggataaa ggagtcgtta tagcggggga catcaccgta
     4381 ggaattgcag atgtcgagct attaacgata aagatccgct tgattgtggc ttcggttgat
     4441 aaggcaaaag aaatcggcat ggactggtgg gaaaatgatc cgtatctcag ttcaaaagga
     4501 gccaataaca aagcgctcga agaagaaaat aaaatgctgc atgagcggtt aaaaacgctt
     4561 gaagaaaaaa tagaaacgaa acgttaaaaa ctgtacgcta cttaaaaaat ggagggattt
     4621 acaatggcaa ctgaaacaaa attagataac acacaggcag aaaacaagga aaataaaaat
     4681 gcggaaaacg gttcaaaaga aaagaacggt tcaaaagcaa gcaaaacaac aagcagcggg
     4741 ccaatcaaac gagcggtagc aggaggcatc atcggtgcaa cgattggata tgtatcgact
     4801 cctgaaaatc gaaaaagtct ccttgaccgc attgatacag acgaattaaa aagcaaagca
     4861 tctgatttag gaacaaaggt aaaagaaaaa tcaaaaagca gcgtggccag cctgaaaaca
     4921 tctgcgggaa gcttgtttaa aaaagataaa gataaatcaa aagatgatga agaaaacgta
     4981 aattcttcta gtagcgaaac agaagacgat aacgttcaag agtacgacga gttaaaagaa
     5041 gaaaatcaaa ctcttcaaga tcgcttatca cagcttgaag aaaaaatgaa catgcttgtt
     5101 gagcttagcc tcaataaaaa tcaagacgaa gaagcggaag atacagattc cgacgaagaa
     5161 gagaacgatg agaacgatga aaacgatgaa aacgagcagg acgatgaaaa cgaagaagaa
     5221 acatctaagc cacgtaaaaa ggataaaaaa gaagctgagg aagaagaaag tgaaagtgac
     5281 gaagacagcg aggaagaaga ggaagattct cgctcaaaca aaaaaaataa aaaagtaaaa
     5341 acagaagaag aagacgaaga tgaaagcgaa gaagaaaaaa aggaagcgaa accaaaaaag
     5401 tcaacagcta aaaaatcaaa aaatacaaaa gcaaagaaaa acacggacga agaagatgat
     5461 gaagcaacat ctctttctag tgaagacgat acaacagcct aagacgtcgt cacatttgct
     5521 tctgcttgct gcagtccgga ttcgcgaacg cgtctcgaga gatccggctg ctaacaaagc
     5581 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     5641 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccgactagta
     5701 gcccgcctaa tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat
     5761 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5821 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5881 tgccttcctg tttttgctca cccagaaacg ctcgtgaaag taaaagacgc agaggaccaa
     5941 ttgggggcac gagtgggata catagaactg gacttgaata gcggtaaaat ccttgagagt
     6001 tttcgccctg aagagcgttt tccaatgatg agcactttca aagttctgct atgtggagca
     6061 gtattatccc gtgtagatgc ggggcaagag caactcggac gacgaataca ctattcgcag
     6121 aatgacttgg ttgaatactc cccagtgaca gaaaagcacc ttacggacgg aatgacggta
     6181 agagaattat gtagtgccgc cataacgatg agtgataaca ctgcggcgaa cttacttctg
     6241 acaaccatcg gtggaccgaa ggaattaacc gcttttttgc acaatatggg agaccatgta
     6301 actcgccttg accgttggga accagaactg aatgaagcca taccaaacga cgagcgagac
     6361 accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg cgaactactt
     6421 actctggctt cacggcaaca attaatagac tggcttgaag cggataaagt tgcaggacca
     6481 ctactgcgtt cggcacttcc tgctggctgg tttattgctg ataaatctgg ggcaggagag
     6541 cgtggttcac ggggtatcat tgccgcactt ggaccagatg gtaagccttc ccgtatcgta
     6601 gttatctaca cgacgggtag tcaggcaact atggacgaac gaaatagaca gattgctgaa
     6661 ataggggctt cactgattaa gcattggtaa accgatacaa ttaaaggctc cttttggagc
     6721 cttttttttt ggacggaccg gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6901 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7321 gatgctcgtc aggggggcgg agcctatgga aa
//
