LOCUS       Exported                7481 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 7481)
  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..7481
                     /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=GS.fix_rev
     primer_bind     complement(3699..3720)
                     /label=gvpG_rev
     primer_bind     3732..3766
                     /label=GS.fix_fwd
     primer_bind     3743..3757
                     /label=gvpS_fwd
     CDS             3749..4036
                     /codon_start=1
                     /label=GvpS
                     /translation="MSLKQSMENKDIALIDILDVILDKGVAIKGDLIISIAGVDLVYLD
                     LRVLISSVETLVQAKEGNHKPITSEQFDKQKEELMDATGQPSKWTNPLGS"
     primer_bind     complement(4052..4071)
                     /label=gvpS_rev
     misc_feature    4071^4072
                     /label=SK
     primer_bind     4072..4087
                     /label=gvpK_fwd
     misc_feature    4093..4101
                     /label=RBS
     CDS             4108..4392
                     /codon_start=1
                     /label=gvpK
                     /translation="MQPVSQANGRIHLDPDQAEQGLAQLVMTVIELLRQIVERHAMRRV
                     EGGTLTDEQIENLGIALMNLEEKMDELKEVFGLDAEDLNIDLGPLGSLL"
     primer_bind     complement(4383..4398)
                     /label=gvpK_rev
     misc_feature    4398^4399
                     /label=KJ
     primer_bind     4399..4417
                     /label=gvpJ_fwd
     misc_feature    4402..4410
                     /label=RBS
     CDS             4414..4716
                     /codon_start=1
                     /label=GvpJ
                     /translation="MAVEHNMQSSTIVDVLEKILDKGVVIAGDITVGIADVELLTIKIR
                     LIVASVDKAKEIGMDWWENDPYLSSKGANNKALEEENKMLHERLKTLEEKIETKR"
     misc_feature    4539
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4545..4566
                     /label=Bm6seqF
     primer_bind     complement(4711..4732)
                     /label=gvpJ_rev
     misc_feature    4732^4733
                     /label=JT
     primer_bind     4733..4756
                     /label=gvpT_fwd
     misc_feature    4739..4747
                     /label=RBS
     CDS             4753..5631
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4774..4800
                     /label=Bm6.1seq
     misc_feature    5077
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    5140
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     5214..5240
                     /label=Bm7seqF
     primer_bind     complement(5619..5636)
                     /label=gvpT_rev
     misc_feature    5636
                     /label=TU
     primer_bind     5637..5654
                     /label=vector_fwd
     misc_feature    5637
                     /label=UV
     terminator      5753..5800
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5867..5958
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5948..5953
                     /label=Emopec_63.3%
     CDS             5959..6819
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6893..7481
                     /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 gaaacattat gtctcttaaa caatccatgg agaataaaga
     3781 tattgctctt attgatattt tagatgtcat tttagataaa ggagtcgcca ttaaaggaga
     3841 cttaatcatt tccatagctg gcgtcgattt agtgtatttg gatttgcggg tgcttatttc
     3901 ttcggttgaa acgcttgtgc aagcaaaaga aggaaatcac aaaccaatca cttctgaaca
     3961 atttgataaa caaaaggaag agttaatgga cgcgaccggc cagccgagca aatggaccaa
     4021 tccgcttggc tcctgaccct tgcgagcagt cgtatgcttt ctcgaattcc gagcggttaa
     4081 gcgtgacagt ccaggaggaa ttaatggatg caaccggtca gccaagcaaa tggacgaatc
     4141 cacttggatc ctgatcaagc tgaacaaggc ttagcgcagc ttgtgatgac agttattgag
     4201 ctattgaggc aaatcgttga acgtcatgcc atgaggcggg tggagggtgg aacgttgacg
     4261 gacgaacaaa ttgaaaactt aggaattgca ctaatgaact tagaagaaaa aatggacgag
     4321 ttgaaagagg tgttcggtct ggatgcagaa gatttaaata ttgatcttgg accgctaggc
     4381 agcctgcttt aagcggtcag taggaggaac agtatggcag tcgaacataa tatgcagtca
     4441 agtacgattg tagatgtgct cgaaaagatt ttggataaag gagtcgttat agcgggggac
     4501 atcaccgtag gaattgcaga tgtcgagcta ttaacgataa agatccgctt gattgtggct
     4561 tcggttgata aggcaaaaga aatcggcatg gactggtggg aaaatgatcc gtatctcagt
     4621 tcaaaaggag ccaataacaa agcgctcgaa gaagaaaata aaatgctgca tgagcggtta
     4681 aaaacgcttg aagaaaaaat agaaacgaaa cgttaaaaac tgtacgctac ttaaaaaatg
     4741 gagggattta caatggcaac tgaaacaaaa ttagataaca cacaggcaga aaacaaggaa
     4801 aataaaaatg cggaaaacgg ttcaaaagaa aagaacggtt caaaagcaag caaaacaaca
     4861 agcagcgggc caatcaaacg agcggtagca ggaggcatca tcggtgcaac gattggatat
     4921 gtatcgactc ctgaaaatcg aaaaagtctc cttgaccgca ttgatacaga cgaattaaaa
     4981 agcaaagcat ctgatttagg aacaaaggta aaagaaaaat caaaaagcag cgtggccagc
     5041 ctgaaaacat ctgcgggaag cttgtttaaa aaagataaag ataaatcaaa agatgatgaa
     5101 gaaaacgtaa attcttctag tagcgaaaca gaagacgata acgttcaaga gtacgacgag
     5161 ttaaaagaag aaaatcaaac tcttcaagat cgcttatcac agcttgaaga aaaaatgaac
     5221 atgcttgttg agcttagcct caataaaaat caagacgaag aagcggaaga tacagattcc
     5281 gacgaagaag agaacgatga gaacgatgaa aacgatgaaa acgagcagga cgatgaaaac
     5341 gaagaagaaa catctaagcc acgtaaaaag gataaaaaag aagctgagga agaagaaagt
     5401 gaaagtgacg aagacagcga ggaagaagag gaagattctc gctcaaacaa aaaaaataaa
     5461 aaagtaaaaa cagaagaaga agacgaagat gaaagcgaag aagaaaaaaa ggaagcgaaa
     5521 ccaaaaaagt caacagctaa aaaatcaaaa aatacaaaag caaagaaaaa cacggacgaa
     5581 gaagatgatg aagcaacatc tctttctagt gaagacgata caacagccta agacgtcgtc
     5641 acatttgctt ctgcttgctg cagtccggat tcgcgaacgc gtctcgagag atccggctgc
     5701 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     5761 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     5821 cgactagtag cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc
     5881 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5941 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6001 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca
     6061 gaggaccaat tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc
     6121 cttgagagtt ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta
     6181 tgtggagcag tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac
     6241 tattcgcaga atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga
     6301 atgacggtaa gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac
     6361 ttacttctga caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga
     6421 gaccatgtaa ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac
     6481 gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc
     6541 gaactactta ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt
     6601 gcaggaccac tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg
     6661 gcaggagagc gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc
     6721 cgtatcgtag ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag
     6781 attgctgaaa taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc
     6841 ttttggagcc tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc
     6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa a
//
