LOCUS       Exported                7547 bp ds-DNA     circular SYN 10-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  AUTHORS   Lu Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7547
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        39..130
                     /gene="bla"
                     /label=AmpR promoter
     RBS             120..125
                     /label=Emopec_63.3%
     CDS             131..991
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1065..1653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1840..1979
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2081..2272)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        2509..2527
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2673..2693)
                     /label=RMpC_RMBB_rev
     primer_bind     complement(2673..2693)
                     /label=RMp2A_RMBB_rev
     primer_bind     2693..2724
                     /label=RMpC_RMBB_fwd
     primer_bind     2694..2724
                     /label=RMp2A_RMBB_fwd
     primer_bind     complement(2728..2753)
                     /label=vector_rev
     misc_feature    2753
                     /label=VB
     primer_bind     2754..2776
                     /label=gvpR_fwd
     misc_feature    2754
                     /label=BR
     primer_bind     2754
                     /label=Bm1seq
     misc_feature    2767..2775
                     /label=RBS
     CDS             2782..3048
                     /codon_start=1
                     /label=GvpR
                     /translation="MEIKKIMQAVNDFFGEHVAPPHKITSVEATEDEGWRVIVEVIEER
                     EYMKKYAKDEMLGTYECFVNKEKEVISFKRLDVRYRSAIGIEA"
     primer_bind     complement(3039..3060)
                     /label=gvpR_rev
     misc_feature    3060^3061
                     /label=RN
     primer_bind     3061..3081
                     /label=gvpN_fwd
     misc_feature    3062..3070
                     /label=RBS
     CDS             3077..4003
                     /codon_start=1
                     /label=GvpN
                     /translation="MTVLTDKRKKGSGAFIQDDETKEVLSRALSYLKSGYSIHFTGPAG
                     GGKTSLARALAKKRKRPVMLMHGNHELNNKDLIGDFTGYTSKKVIDQYVRSVYKKDEQV
                     SENWQDGRLLEAVKNGYTLIYDEFTRSKPATNNIFLSILEEGVLPLYGVKMTDPFVRVH
                     PDFRVIFTSNPAEYAGVYDTQDALLDRLITMFIDYKDIDRETAILTEKTDVEEDEARTI
                     VTLVANVRNRSGDENSSGLSLRASLMIATLATQQDIPIDGSDEDFQTLCIDILHHPLTK
                     CLDEENAKSKAEKIILEECKNIDTEEK"
     primer_bind     3107..3132
                     /label=Bm1.1seqF
     misc_feature    3139
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     3509..3528
                     /label=Bm2seqF
     primer_bind     complement(3968..4024)
                     /label=G dbfG r
     primer_bind     complement(3974..4003)
                     /label=gvpN_rev
     misc_feature    4003
                     /label=NF
     misc_feature    4004
                     /label=FG
     CDS             4008..4052
                     /codon_start=1
                     /label=GvpG
                     /translation="EQWEELTQKRNEES"
     primer_bind     4008..4036
                     /label=G dbfG f
     primer_bind     4008..4025
                     /label=Bm4seqF
     primer_bind     complement(4016..4037)
                     /label=gvpG_rev
     misc_feature    4037^4038
                     /label=GL
     primer_bind     4038..4057
                     /label=gvpL_fwd
     misc_feature    4039..4047
                     /label=RBS
     CDS             4053..4862
                     /codon_start=1
                     /label=GvpL
                     /translation="MGELLYLYGLIPTKEAAAIEPFPSYKGFDGEHSLYPIAFDQVTAV
                     VSKLDADTYSEKVIQEKMEQDMSWLQEKAFHHHETVAALYEEFTIIPLKFCTIYKGEES
                     LQAAIEINKEKIENSLTLLQGNEEWNVKIYCDDTELKKGISETNESVKAKKQEISHLSP
                     GRQFFEKKKIDQLIEKELELHKNKVCEEIHDKLKELSLYDSVKKNWSKDVTGAAEQMAW
                     NSVFLLPSLQITKFVNEIEELQQRLENKGWKFEVTGPWPPYHFSSFA"
     misc_feature    4123
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4192
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4642
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4773..4799
                     /label=Bm5seqF
     primer_bind     complement(4846..4863)
                     /label=gvpL_rev
     misc_feature    4863^4864
                     /label=LS
     primer_bind     4864..4888
                     /label=gvpS_fwd
     misc_feature    4864..4872
                     /label=RBS
     CDS             4880..5167
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(5183..5202)
                     /label=gvpS_rev
     misc_feature    5202^5203
                     /label=SK
     primer_bind     5203..5218
                     /label=gvpK_fwd
     misc_feature    5224..5232
                     /label=RBS
     CDS             5239..5523
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(5514..5529)
                     /label=gvpK_rev
     misc_feature    5529^5530
                     /label=KJ
     primer_bind     5530..5548
                     /label=gvpJ_fwd
     misc_feature    5533..5541
                     /label=RBS
     CDS             5545..5847
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    5670
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     5676..5697
                     /label=Bm6seqF
     primer_bind     complement(5842..5863)
                     /label=gvpJ_rev
     misc_feature    5863^5864
                     /label=JT
     primer_bind     5864..5887
                     /label=gvpT_fwd
     misc_feature    5870..5878
                     /label=RBS
     CDS             5884..6762
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     5905..5931
                     /label=Bm6.1seq
     misc_feature    6208
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    6271
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     6345..6371
                     /label=Bm7seqF
     primer_bind     complement(6750..6767)
                     /label=gvpT_rev
     misc_feature    6767^6768
                     /label=TU
     primer_bind     6768..6788
                     /label=gvpU_fwd
     misc_feature    6770..6778
                     /label=RBS
     CDS             6786..7202
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     7125..7149
                     /label=GvpUseqF
     misc_feature    7355^7356
                     /label=UV
     primer_bind     7356..7373
                     /label=vector_fwd
     terminator      7472..7519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg cagaggacca
      241 attgggggca cgagtgggat acatagaact ggacttgaat agcggtaaaa tccttgagag
      301 ttttcgccct gaagagcgtt ttccaatgat gagcactttc aaagttctgc tatgtggagc
      361 agtattatcc cgtgtagatg cggggcaaga gcaactcgga cgacgaatac actattcgca
      421 gaatgacttg gttgaatact ccccagtgac agaaaagcac cttacggacg gaatgacggt
      481 aagagaatta tgtagtgccg ccataacgat gagtgataac actgcggcga acttacttct
      541 gacaaccatc ggtggaccga aggaattaac cgcttttttg cacaatatgg gagaccatgt
      601 aactcgcctt gaccgttggg aaccagaact gaatgaagcc ataccaaacg acgagcgaga
      661 caccacaatg cctgcggcaa tggcaacaac attacgcaaa ctattaactg gcgaactact
      721 tactctggct tcacggcaac aattaataga ctggcttgaa gcggataaag ttgcaggacc
      781 actactgcgt tcggcacttc ctgctggctg gtttattgct gataaatctg gggcaggaga
      841 gcgtggttca cggggtatca ttgccgcact tggaccagat ggtaagcctt cccgtatcgt
      901 agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga
      961 aataggggct tcactgatta agcattggta aaccgataca attaaaggct ccttttggag
     1021 cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt
     1081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1141 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1201 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1261 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1321 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1381 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1441 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1501 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1561 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1621 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc cctttttacg
     1681 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1741 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1801 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     1861 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     1921 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     1981 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2041 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2101 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     2161 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     2221 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     2281 ccgtgtaagg gggatttctg ttcatggggt aatgataccg atgaaacgag agaggatgct
     2341 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2401 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2461 cgttaataca gatgtaggtg ttggcaccat gcatcgatcc cgcgaaatta atacgactca
     2521 ctatagggag accacaacgg tttccctcta gagatagctt agcgatgtga aagaagctgg
     2581 agaaaacttt caagaaaaag tttcttcttc agacgataaa gagaagtcaa aaaataagcg
     2641 gaaaataaaa ggcgtaaatc agattaaaaa agcaacggat attaaaagct ctactaagat
     2701 taaaagttct aatgatatta aatcatctac agatttaaaa acaatgggat cattagatct
     2761 aactattgga ggctactaaa aatggaaatt aaaaaaatta tgcaagccgt gaacgacttt
     2821 ttcggtgaac acgtagctcc tcctcataaa attacctcgg tggaagctac tgaagatgaa
     2881 ggttggagag ttattgttga agtcattgaa gaacgagaat atatgaaaaa atacgccaaa
     2941 gatgaaatgc tcggaacgta cgagtgcttt gtaaataaag aaaaagaagt catttcattc
     3001 aaacgactcg acgtcagata tagaagcgcc attggcattg aagcataaac gataagatgg
     3061 caggaggaac gtaaaaatga ccgtcttaac agacaaaagg aaaaaaggca gtggagcttt
     3121 tatacaagat gacgagacaa aagaggttct ttcaagagcg ctgagctatt taaaatccgg
     3181 ctattccatt cattttacag gtcctgccgg cggaggcaaa acctctttag cgcgagcgct
     3241 tgctaaaaag agaaagcgtc ctgtaatgct gatgcacggg aatcacgagc tcaacaacaa
     3301 agatttaatt ggcgatttta cgggatacac gagcaaaaaa gtaatcgacc agtacgttcg
     3361 ttctgtctat aaaaaagatg aacaggtgag tgaaaactgg caggatggcc gattgcttga
     3421 agctgtaaaa aatggctata cgctgattta cgacgaattt actcgttcta agcctgcgac
     3481 gaataatatc tttctatcga tattagaaga aggcgtgctg ccgctgtatg gagtaaaaat
     3541 gaccgatcct tttgtgcgcg tgcatcccga tttccgcgtc atcttcacaa gcaatccagc
     3601 tgagtatgcc ggcgtatatg atacgcaaga tgcgcttctc gacaggttaa ttaccatgtt
     3661 tattgattat aaagacatcg acagagagac agcgatttta acggagaaaa cggacgtaga
     3721 agaagatgaa gcgcgcacaa ttgtaacgct cgtagcaaac gtgcgaaacc gctctggaga
     3781 cgaaaacagc agcggactta gcctgcgggc ttcgcttatg atcgctaccc ttgccacgca
     3841 gcaagacatt cctatcgatg gaagtgacga agattttcaa acgttatgta tcgatatttt
     3901 gcatcatccg cttaccaaat gtttggatga agaaaatgca aaaagcaaag ccgaaaaaat
     3961 cattttagaa gaatgtaaga atatagacac tgaagaaaag taagtaggaa caatgggaag
     4021 agctaacaca aaaaagaaat gaggaatcct agatgggaga attactgtat ttatacggtt
     4081 taattccaac aaaagaagca gcagccatag agccgtttcc atcttataag gggtttgacg
     4141 gagaacattc actgtaccca attgcgtttg atcaggtgac ggctgtagtt tctaagctgg
     4201 atgctgacac ctattcagaa aaagtgattc aagaaaaaat ggagcaggat atgagctggc
     4261 tgcaggaaaa agcatttcat catcacgaaa cggtagccgc tttgtacgaa gaatttacga
     4321 tcattccatt aaaattttgc accatttata aaggtgaaga aagtctgcag gcagctattg
     4381 agattaacaa agaaaagata gagaattcac tgacgctgct tcaaggaaat gaagagtgga
     4441 atgtgaaaat ttactgtgat gatacagagc ttaaaaaagg aatcagcgaa acgaatgaaa
     4501 gcgtgaaagc gaaaaaacaa gaaattagtc acttatcacc aggaagacag ttttttgaaa
     4561 agaaaaaaat agatcagctg attgaaaaag aattagagct tcacaaaaac aaagtgtgtg
     4621 aagagataca tgacaagcta aaagaattat cgctttatga ctctgttaaa aagaattgga
     4681 gcaaagacgt aactggcgca gctgaacaga tggcgtggaa cagcgtgttt cttctcccgt
     4741 ctctgcagat tactaagttc gtaaacgaaa tagaagagct tcagcaaagg cttgaaaata
     4801 aaggctggaa gtttgaagtg acgggaccat ggccgcccta tcatttctcg agctttgcgt
     4861 aaagtgagga attaacatta tgtctcttaa acaatccatg gagaataaag atattgctct
     4921 tattgatatt ttagatgtca ttttagataa aggagtcgcc attaaaggag acttaatcat
     4981 ttccatagct ggcgtcgatt tagtgtattt ggatttgcgg gtgcttattt cttcggttga
     5041 aacgcttgtg caagcaaaag aaggaaatca caaaccaatc acttctgaac aatttgataa
     5101 acaaaaggaa gagttaatgg acgcgaccgg ccagccgagc aaatggacca atccgcttgg
     5161 ctcctgaccc ttgcgagcag tcgtatgctt tctcgaattc cgagcggtta agcgtgacag
     5221 tccaggagga attaatggat gcaaccggtc agccaagcaa atggacgaat ccacttggat
     5281 cctgatcaag ctgaacaagg cttagcgcag cttgtgatga cagttattga gctattgagg
     5341 caaatcgttg aacgtcatgc catgaggcgg gtggagggtg gaacgttgac ggacgaacaa
     5401 attgaaaact taggaattgc actaatgaac ttagaagaaa aaatggacga gttgaaagag
     5461 gtgttcggtc tggatgcaga agatttaaat attgatcttg gaccgctagg cagcctgctt
     5521 taagcggtca gtaggaggaa cagtatggca gtcgaacata atatgcagtc aagtacgatt
     5581 gtagatgtgc tcgaaaagat tttggataaa ggagtcgtta tagcggggga catcaccgta
     5641 ggaattgcag atgtcgagct attaacgata aagatccgct tgattgtggc ttcggttgat
     5701 aaggcaaaag aaatcggcat ggactggtgg gaaaatgatc cgtatctcag ttcaaaagga
     5761 gccaataaca aagcgctcga agaagaaaat aaaatgctgc atgagcggtt aaaaacgctt
     5821 gaagaaaaaa tagaaacgaa acgttaaaaa ctgtacgcta cttaaaaaat ggagggattt
     5881 acaatggcaa ctgaaacaaa attagataac acacaggcag aaaacaagga aaataaaaat
     5941 gcggaaaacg gttcaaaaga aaagaacggt tcaaaagcaa gcaaaacaac aagcagcggg
     6001 ccaatcaaac gagcggtagc aggaggcatc atcggtgcaa cgattggata tgtatcgact
     6061 cctgaaaatc gaaaaagtct ccttgaccgc attgatacag acgaattaaa aagcaaagca
     6121 tctgatttag gaacaaaggt aaaagaaaaa tcaaaaagca gcgtggccag cctgaaaaca
     6181 tctgcgggaa gcttgtttaa aaaagataaa gataaatcaa aagatgatga agaaaacgta
     6241 aattcttcta gtagcgaaac agaagacgat aacgttcaag agtacgacga gttaaaagaa
     6301 gaaaatcaaa ctcttcaaga tcgcttatca cagcttgaag aaaaaatgaa catgcttgtt
     6361 gagcttagcc tcaataaaaa tcaagacgaa gaagcggaag atacagattc cgacgaagaa
     6421 gagaacgatg agaacgatga aaacgatgaa aacgagcagg acgatgaaaa cgaagaagaa
     6481 acatctaagc cacgtaaaaa ggataaaaaa gaagctgagg aagaagaaag tgaaagtgac
     6541 gaagacagcg aggaagaaga ggaagattct cgctcaaaca aaaaaaataa aaaagtaaaa
     6601 acagaagaag aagacgaaga tgaaagcgaa gaagaaaaaa aggaagcgaa accaaaaaag
     6661 tcaacagcta aaaaatcaaa aaatacaaaa gcaaagaaaa acacggacga agaagatgat
     6721 gaagcaacat ctctttctag tgaagacgat acaacagcct aagacgtaaa ggaggaaaga
     6781 aagacatgag tacaggccct tctttttcaa ctaaagacaa tacgcttgaa tactttgtga
     6841 aagcttctaa taaacacggc ttttcacttg atatttcatt aaatgtaaac ggcgctgtga
     6901 tttccggtac catgatttca gcaaaagaat actttgatta cttaagcgaa acgtttgaag
     6961 aaggcagtga agtggctcag gcgctaagcg aacaattctc tttagcaagc gaagcgagcg
     7021 aatcaaacgg agaagcagaa gcccatttta ttcatttgaa aaatacaaag atttactgtg
     7081 gagacagtaa atctactcct tctaaaggca aaatcttttg gagagggaaa atagcagaag
     7141 tagacgggtt tttcttagga aagatttctg atgcaaaatc aacgagtaaa aagagttcat
     7201 aaaaaacggc ggggtgattg ccccgccgtt ttttagtgat gtgatgagat gtgcagcttc
     7261 ttttttcgat acatgagcgg agtcatcttc atttcttttc gaaactgatt aataaaatag
     7321 cttgtactat taaaccctac ttgataggcg acttccgtca catttgcttc tgcttgctgc
     7381 agtccggatt cgcgaacgcg tctcgagaga tccggctgct aacaaagccc gaaaggaagc
     7441 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     7501 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc gactagt
//
