LOCUS       sequence-52225-e        5713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5713)
  AUTHORS   MJFF
  TITLE     Michael J Fox Foundation plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5713)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pEMB12/LRRK2_BC_1847_2170_P1865CCC
FEATURES             Location/Qualifiers
     source          1..5713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        615..899
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     822..841
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             937..954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1285..1302
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2317..2334)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2487..2573
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2665..2692
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2711..2802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2803..2871,2872..3663)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3021..3040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3705..4160
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3792..3811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4271..4859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4760..4779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5013..5030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5196..5218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(5423..5713,1..588))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
       61 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
      121 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
      181 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
      241 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
      301 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
      361 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
      421 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
      481 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
      541 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
      601 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
      661 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
      721 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
      781 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
      841 cataagatta gcggctccta cctgacgctt tttatcgcaa ctctctactg tttctccata
      901 cccgtttttt tgggctagca ggaggaattc atgaagcatc accaccatca ccatctcgag
      961 tccgataaga taatccacct gactgacgac tccttcgata ccgacgtgct gaaggcagac
     1021 ggtgctatcc tcgttgactt ctgggcccac tggtgtggtc cttgcaagat gatagctcct
     1081 attcttgacg agatcgcaga cgaataccaa ggtaagctga ccgttgctaa gctgaacatt
     1141 gatcataacc ctggcaccgc acctaaatac ggcatccgtg gcatcccaac tttgctcctg
     1201 ttcaagaatg gtgaagtagc agctaccaag gtaggcgcac tgtctaaagg ccagctgaaa
     1261 gaatttctgg acgccaactt agctttagtt cctcgcggat ccaggctgac catccccatc
     1321 tcccagattg cacccgacct tatcctcgct gacctgcccc gcaacatcat gctgaacaat
     1381 gacgagctgg agtttgaaca agcacccgag ttcctgctag gcgacggctc tttcggttca
     1441 gtttaccgtg ctgcttacga gggtgaagag gttgccgtga agatttttaa caagcacact
     1501 tcgctccgtc tgctgcgtca ggagcttgtg gttctctgcc acctgcacca cccttccctg
     1561 atctccctgc tggccgccgg cattaggcct cgcatgctag tgatggaact ggccagtaag
     1621 ggttcattgg accgcctgct gcagcaggac aaggctagcc tgacccgtac gctccagcac
     1681 cgtatcgccc tgcatgtggc cgacggactg cgctacttgc acagtgccat gatcatttat
     1741 cgcgatctga agcctcataa tgttcttctg ttcacgctct acccaaacgc agctatcatt
     1801 gccaaaattg ccgattacgg catagcccag tactgctgtc gtatgggtat caagactagc
     1861 gagggtacac ctgggttccg cgctcccgaa gtcgcacgcg ggaacgtgat ctacaaccag
     1921 caggccgacg tgtactcgtt tggcctgctc ctgtatgaca ttctcactac cggaggccgg
     1981 attgtggagg gcctgaaatt tcccaacgag ttcgacgagc tggagatcca gggcaagctg
     2041 cctgaccccg tgaaggagta cggttgtgca ccctggccta tggtggagaa gctcattaag
     2101 cagtgtctga aggagaaccc tcaggaacgc cccacaagcg cccaggtgtt cgacattcta
     2161 aactctgcag agttagtgtg tctgactagg cgtatcttac tcccgaagaa cgtgatcgtt
     2221 gagtgcatgg tggcaacgca ccacaactcg cgtaacgcta gcatctggtt aggatgataa
     2281 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     2341 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     2401 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     2461 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2521 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     2581 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     2641 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     2701 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2761 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2821 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2881 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2941 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3001 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3061 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3481 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3661 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     3721 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3781 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3841 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3901 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3961 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4021 ttgaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4081 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4141 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4201 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4261 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4321 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4381 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4441 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4501 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4561 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4621 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4681 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4741 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4801 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4861 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4921 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4981 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5041 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt
     5101 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5161 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5221 ttcaccgtca tcaccgaaac gcgcgaggca gcaaggagat ggcgcgccgg tgatgccggc
     5281 cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat
     5341 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt
     5401 caattgtctg attcgttacc aattatgaca acttgacggc tacatcattc actttttctt
     5461 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     5521 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     5581 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     5641 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     5701 cgacgctggc gat
//
