LOCUS       Exported                7376 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 7376)
  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..7376
                     /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    2685^2686
                     /label=NF
     primer_bind     2686..2705
                     /label=gvpF_fwd
     misc_feature    2686..2694
                     /label=RBS
     CDS             2700..3467
                     /codon_start=1
                     /label=GvpF
                     /translation="MSETNETGIYIFSAIQTDKDEEFGAVEVEGTKAETFLIRYKDAAM
                     VAAEVPMKIYHPNRQNLLMHQNAVAAIMDKNDTVIPISFGNVFKSKEDVKVLLENLYPQ
                     FEKLFPAIKGKIEVGLKVIGKKEWLEKKVNENPELEKVSASVKGKSEAAGYYERIQLGG
                     MAQKMFTSLQKEVKTDVFSPLEEAAEAAKANEPTGETMLLNASFLINREDEAKFDEKVN
                     EAHENWKDKADFHYSGPWPAYNFVNIRLKVEEK"
     primer_bind     2933..2960
                     /label=Bm3seqF
     primer_bind     complement(3432..3452)
                     /label=gvpF_rev
     misc_feature    3452^3453
                     /label=FG
     primer_bind     3453..3475
                     /label=gvpG_fwd
     misc_feature    3457..3465
                     /label=RBS
     CDS             3469..3720
                     /codon_start=1
                     /label=GvpG
                     /translation="VLHKLVTAPINLVVKIGEKVQEEADKQLYDLPTIQQKLIQLQMMF
                     ELGEIPEEAFQEKEDELLMRYEIAKRREIEQWEELTQKR"
     primer_bind     3681..3708
                     /label=Bm4seqF
     primer_bind     complement(3699..3731)
                     /label=GK.fix_rev
     primer_bind     complement(3699..3720)
                     /label=gvpG_rev
     misc_feature    3720
                     /label=GL
     primer_bind     3732..3758
                     /label=GK.fix_fwd
     CDS             3749..4033
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4024..4039)
                     /label=gvpK_rev
     misc_feature    4039
                     /label=KJ
     primer_bind     4040..4063
                     /label=gvpT_fwd
     misc_feature    4040
                     /label=JT
     misc_feature    4046..4054
                     /label=RBS
     CDS             4060..4938
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4081..4107
                     /label=Bm6.1seq
     misc_feature    4384
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4447
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4521..4547
                     /label=Bm7seqF
     primer_bind     complement(4926..4943)
                     /label=gvpT_rev
     misc_feature    4943^4944
                     /label=TU
     primer_bind     4944..4964
                     /label=gvpU_fwd
     misc_feature    4946..4954
                     /label=RBS
     CDS             4962..5378
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5301..5325
                     /label=GvpUseqF
     misc_feature    5531^5532
                     /label=UV
     primer_bind     5532..5549
                     /label=vector_fwd
     terminator      5648..5695
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5762..5853
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5843..5848
                     /label=Emopec_63.3%
     CDS             5854..6714
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6788..7376
                     /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 agtaaaggag cttgaaaaca
     2701 tgagtgaaac aaacgaaaca ggtatttata tttttagcgc cattcaaacg gataaagacg
     2761 aagaatttgg cgccgtggaa gtagaaggaa caaaagctga aacatttttg attcgctaca
     2821 aagacgcggc tatggtagca gctgaagtac cgatgaaaat ttatcatcct aatcgccaaa
     2881 atttattaat gcatcaaaac gcagtagcag cgattatgga caagaacgat acggttattc
     2941 caatcagctt tgggaatgta ttcaaatcaa aagaagacgt aaaagttctt ttggaaaacc
     3001 tttatccgca gtttgaaaag ctgtttccag cgatcaaagg aaaaattgaa gtcggtttaa
     3061 aagtaattgg gaaaaaagaa tggcttgaga aaaaagtaaa cgaaaatcct gaacttgaga
     3121 aagtatcagc atccgtaaaa ggaaaatcag aagcagccgg ttattatgag cgtattcaac
     3181 ttggaggaat ggctcaaaag atgtttactt ccctgcaaaa agaagtcaag acagatgtgt
     3241 tttctccgct tgaagaagca gcggaagcag caaaagcaaa tgagccaacg ggcgaaacga
     3301 tgcttttaaa cgcgtctttc ttaattaacc gagaagatga agcgaagttt gatgaaaaag
     3361 taaatgaagc gcatgaaaac tggaaagaca aagccgattt tcattacagc ggtccttggc
     3421 ctgcttataa ttttgtgaac attcgcctaa aagtagaaga gaaataacgt gcttcacaaa
     3481 ttagtaaccg cacccattaa ccttgtagtg aaaatcggcg aaaaagtaca ggaagaagct
     3541 gataaacagc tatatgacct tccgacgatt cagcaaaagc tcattcagct tcaaatgatg
     3601 tttgagcttg gtgaaattcc agaagaagcg tttcaagaaa aagaagatga attgttaatg
     3661 aggtacgaaa ttgcgaaacg cagagaaatt gaacaatggg aagagctaac acaaaaaaga
     3721 aatgaagaat cctaatagga gataatggat gcaaccggtc agccaagcaa atggacgaat
     3781 ccacttggat cctgatcaag ctgaacaagg cttagcgcag cttgtgatga cagttattga
     3841 gctattgagg caaatcgttg aacgtcatgc catgaggcgg gtggagggtg gaacgttgac
     3901 ggacgaacaa attgaaaact taggaattgc actaatgaac ttagaagaaa aaatggacga
     3961 gttgaaagag gtgttcggtc tggatgcaga agatttaaat attgatcttg gaccgctagg
     4021 cagcctgctt taagcggtca aaaaatggag ggatttacaa tggcaactga aacaaaatta
     4081 gataacacac aggcagaaaa caaggaaaat aaaaatgcgg aaaacggttc aaaagaaaag
     4141 aacggttcaa aagcaagcaa aacaacaagc agcgggccaa tcaaacgagc ggtagcagga
     4201 ggcatcatcg gtgcaacgat tggatatgta tcgactcctg aaaatcgaaa aagtctcctt
     4261 gaccgcattg atacagacga attaaaaagc aaagcatctg atttaggaac aaaggtaaaa
     4321 gaaaaatcaa aaagcagcgt ggccagcctg aaaacatctg cgggaagctt gtttaaaaaa
     4381 gataaagata aatcaaaaga tgatgaagaa aacgtaaatt cttctagtag cgaaacagaa
     4441 gacgataacg ttcaagagta cgacgagtta aaagaagaaa atcaaactct tcaagatcgc
     4501 ttatcacagc ttgaagaaaa aatgaacatg cttgttgagc ttagcctcaa taaaaatcaa
     4561 gacgaagaag cggaagatac agattccgac gaagaagaga acgatgagaa cgatgaaaac
     4621 gatgaaaacg agcaggacga tgaaaacgaa gaagaaacat ctaagccacg taaaaaggat
     4681 aaaaaagaag ctgaggaaga agaaagtgaa agtgacgaag acagcgagga agaagaggaa
     4741 gattctcgct caaacaaaaa aaataaaaaa gtaaaaacag aagaagaaga cgaagatgaa
     4801 agcgaagaag aaaaaaagga agcgaaacca aaaaagtcaa cagctaaaaa atcaaaaaat
     4861 acaaaagcaa agaaaaacac ggacgaagaa gatgatgaag caacatctct ttctagtgaa
     4921 gacgatacaa cagcctaaga cgtaaaggag gaaagaaaga catgagtaca ggcccttctt
     4981 tttcaactaa agacaatacg cttgaatact ttgtgaaagc ttctaataaa cacggctttt
     5041 cacttgatat ttcattaaat gtaaacggcg ctgtgatttc cggtaccatg atttcagcaa
     5101 aagaatactt tgattactta agcgaaacgt ttgaagaagg cagtgaagtg gctcaggcgc
     5161 taagcgaaca attctcttta gcaagcgaag cgagcgaatc aaacggagaa gcagaagccc
     5221 attttattca tttgaaaaat acaaagattt actgtggaga cagtaaatct actccttcta
     5281 aaggcaaaat cttttggaga gggaaaatag cagaagtaga cgggtttttc ttaggaaaga
     5341 tttctgatgc aaaatcaacg agtaaaaaga gttcataaaa aacggcgggg tgattgcccc
     5401 gccgtttttt agtgatgtga tgagatgtgc agcttctttt ttcgatacat gagcggagtc
     5461 atcttcattt cttttcgaaa ctgattaata aaatagcttg tactattaaa ccctacttga
     5521 taggcgactt ccgtcacatt tgcttctgct tgctgcagtc cggattcgcg aacgcgtctc
     5581 gagagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     5641 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     5701 aggaggaact atatccgact agtagcccgc ctaatgagcg ggcttttttt taattcccct
     5761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5821 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctcgtg
     5941 aaagtaaaag acgcagagga ccaattgggg gcacgagtgg gatacataga actggacttg
     6001 aatagcggta aaatccttga gagttttcgc cctgaagagc gttttccaat gatgagcact
     6061 ttcaaagttc tgctatgtgg agcagtatta tcccgtgtag atgcggggca agagcaactc
     6121 ggacgacgaa tacactattc gcagaatgac ttggttgaat actccccagt gacagaaaag
     6181 caccttacgg acggaatgac ggtaagagaa ttatgtagtg ccgccataac gatgagtgat
     6241 aacactgcgg cgaacttact tctgacaacc atcggtggac cgaaggaatt aaccgctttt
     6301 ttgcacaata tgggagacca tgtaactcgc cttgaccgtt gggaaccaga actgaatgaa
     6361 gccataccaa acgacgagcg agacaccaca atgcctgcgg caatggcaac aacattacgc
     6421 aaactattaa ctggcgaact acttactctg gcttcacggc aacaattaat agactggctt
     6481 gaagcggata aagttgcagg accactactg cgttcggcac ttcctgctgg ctggtttatt
     6541 gctgataaat ctggggcagg agagcgtggt tcacggggta tcattgccgc acttggacca
     6601 gatggtaagc cttcccgtat cgtagttatc tacacgacgg gtagtcaggc aactatggac
     6661 gaacgaaata gacagattgc tgaaataggg gcttcactga ttaagcattg gtaaaccgat
     6721 acaattaaag gctccttttg gagccttttt ttttggacgg accggtagaa aagatcaaag
     6781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaa
//
