LOCUS       Exported                7428 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 7428)
  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..7428
                     /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
                     /label=KJ
     primer_bind     4092..4115
                     /label=gvpT_fwd
     misc_feature    4092
                     /label=JT
     misc_feature    4098..4106
                     /label=RBS
     CDS             4112..4990
                     /codon_start=1
                     /label=GvpT
                     /translation="MATETKLDNTQAENKENKNAENGSKEKNGSKASKTTSSGPIKRAV
                     AGGIIGATIGYVSTPENRKSLLDRIDTDELKSKASDLGTKVKEKSKSSVASLKTSAGSL
                     FKKDKDKSKDDEENVNSSSSETEDDNVQEYDELKEENQTLQDRLSQLEEKMNMLVELSL
                     NKNQDEEAEDTDSDEEENDENDENDENEQDDENEEETSKPRKKDKKEAEEEESESDEDS
                     EEEEEDSRSNKKNKKVKTEEEDEDESEEEKKEAKPKKSTAKKSKNTKAKKNTDEEDDEA
                     TSLSSEDDTTA"
     primer_bind     4133..4159
                     /label=Bm6.1seq
     misc_feature    4436
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     misc_feature    4499
                     /label=SNP
                     /note="Bacillus megaterium QM B1551 SNP"
     primer_bind     4573..4599
                     /label=Bm7seqF
     primer_bind     complement(4978..4995)
                     /label=gvpT_rev
     misc_feature    4995^4996
                     /label=TU
     primer_bind     4996..5016
                     /label=gvpU_fwd
     misc_feature    4998..5006
                     /label=RBS
     CDS             5014..5430
                     /codon_start=1
                     /label=GvpU
                     /translation="MSTGPSFSTKDNTLEYFVKASNKHGFSLDISLNVNGAVISGTMIS
                     AKEYFDYLSETFEEGSEVAQALSEQFSLASEASESNGEAEAHFIHLKNTKIYCGDSKST
                     PSKGKIFWRGKIAEVDGFFLGKISDAKSTSKKSS"
     primer_bind     5353..5377
                     /label=GvpUseqF
     misc_feature    5583^5584
                     /label=UV
     primer_bind     5584..5601
                     /label=vector_fwd
     terminator      5700..5747
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5814..5905
                     /gene="bla"
                     /label=AmpR promoter
     RBS             5895..5900
                     /label=Emopec_63.3%
     CDS             5906..6766
                     /codon_start=1
                     /label=Beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWLEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6840..7428
                     /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 caaaaaatgg agggatttac aatggcaact gaaacaaaat tagataacac
     4141 acaggcagaa aacaaggaaa ataaaaatgc ggaaaacggt tcaaaagaaa agaacggttc
     4201 aaaagcaagc aaaacaacaa gcagcgggcc aatcaaacga gcggtagcag gaggcatcat
     4261 cggtgcaacg attggatatg tatcgactcc tgaaaatcga aaaagtctcc ttgaccgcat
     4321 tgatacagac gaattaaaaa gcaaagcatc tgatttagga acaaaggtaa aagaaaaatc
     4381 aaaaagcagc gtggccagcc tgaaaacatc tgcgggaagc ttgtttaaaa aagataaaga
     4441 taaatcaaaa gatgatgaag aaaacgtaaa ttcttctagt agcgaaacag aagacgataa
     4501 cgttcaagag tacgacgagt taaaagaaga aaatcaaact cttcaagatc gcttatcaca
     4561 gcttgaagaa aaaatgaaca tgcttgttga gcttagcctc aataaaaatc aagacgaaga
     4621 agcggaagat acagattccg acgaagaaga gaacgatgag aacgatgaaa acgatgaaaa
     4681 cgagcaggac gatgaaaacg aagaagaaac atctaagcca cgtaaaaagg ataaaaaaga
     4741 agctgaggaa gaagaaagtg aaagtgacga agacagcgag gaagaagagg aagattctcg
     4801 ctcaaacaaa aaaaataaaa aagtaaaaac agaagaagaa gacgaagatg aaagcgaaga
     4861 agaaaaaaag gaagcgaaac caaaaaagtc aacagctaaa aaatcaaaaa atacaaaagc
     4921 aaagaaaaac acggacgaag aagatgatga agcaacatct ctttctagtg aagacgatac
     4981 aacagcctaa gacgtaaagg aggaaagaaa gacatgagta caggcccttc tttttcaact
     5041 aaagacaata cgcttgaata ctttgtgaaa gcttctaata aacacggctt ttcacttgat
     5101 atttcattaa atgtaaacgg cgctgtgatt tccggtacca tgatttcagc aaaagaatac
     5161 tttgattact taagcgaaac gtttgaagaa ggcagtgaag tggctcaggc gctaagcgaa
     5221 caattctctt tagcaagcga agcgagcgaa tcaaacggag aagcagaagc ccattttatt
     5281 catttgaaaa atacaaagat ttactgtgga gacagtaaat ctactccttc taaaggcaaa
     5341 atcttttgga gagggaaaat agcagaagta gacgggtttt tcttaggaaa gatttctgat
     5401 gcaaaatcaa cgagtaaaaa gagttcataa aaaacggcgg ggtgattgcc ccgccgtttt
     5461 ttagtgatgt gatgagatgt gcagcttctt ttttcgatac atgagcggag tcatcttcat
     5521 ttcttttcga aactgattaa taaaatagct tgtactatta aaccctactt gataggcgac
     5581 ttccgtcaca tttgcttctg cttgctgcag tccggattcg cgaacgcgtc tcgagagatc
     5641 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     5701 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     5761 ctatatccga ctagtagccc gcctaatgag cgggcttttt tttaattccc ctatttgttt
     5821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5881 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5941 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctcg tgaaagtaaa
     6001 agacgcagag gaccaattgg gggcacgagt gggatacata gaactggact tgaatagcgg
     6061 taaaatcctt gagagttttc gccctgaaga gcgttttcca atgatgagca ctttcaaagt
     6121 tctgctatgt ggagcagtat tatcccgtgt agatgcgggg caagagcaac tcggacgacg
     6181 aatacactat tcgcagaatg acttggttga atactcccca gtgacagaaa agcaccttac
     6241 ggacggaatg acggtaagag aattatgtag tgccgccata acgatgagtg ataacactgc
     6301 ggcgaactta cttctgacaa ccatcggtgg accgaaggaa ttaaccgctt ttttgcacaa
     6361 tatgggagac catgtaactc gccttgaccg ttgggaacca gaactgaatg aagccatacc
     6421 aaacgacgag cgagacacca caatgcctgc ggcaatggca acaacattac gcaaactatt
     6481 aactggcgaa ctacttactc tggcttcacg gcaacaatta atagactggc ttgaagcgga
     6541 taaagttgca ggaccactac tgcgttcggc acttcctgct ggctggttta ttgctgataa
     6601 atctggggca ggagagcgtg gttcacgggg tatcattgcc gcacttggac cagatggtaa
     6661 gccttcccgt atcgtagtta tctacacgac gggtagtcag gcaactatgg acgaacgaaa
     6721 tagacagatt gctgaaatag gggcttcact gattaagcat tggtaaaccg atacaattaa
     6781 aggctccttt tggagccttt ttttttggac ggaccggtag aaaagatcaa aggatcttct
     6841 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6901 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6961 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     7021 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7081 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7141 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7201 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7261 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7321 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7381 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaa
//
