LOCUS       sequence-256986-        5174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of human DJ1 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5174)
  TITLE     MJFF plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5174)
  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            "Unpublished"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: pET3a-DJ1
FEATURES             Location/Qualifiers
     source          1..5174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(766..784)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      780..827
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1193..1198,1199..1215,1216..1221))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1248..1266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1334..1438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1439..1507,1508..2299)
                     /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(1657..1676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2470..3058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2959..2978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3212..3229
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3244..3383
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3309..3328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3469..3491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3485..3676)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5134..5153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
       61 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatggcaag caagcgcgca ctggtgattc tggccaaggg
      181 cgccgaagag atggaaaccg tgatcccggt tgatgtgatg cgccgcgccg gtattaaagt
      241 gaccgtggca ggtctggccg gcaaagatcc ggtgcagtgc agccgcgatg tggtgatttg
      301 tccggatgcc agcctggaag atgccaaaaa agaggggccg tatgatgtgg tggttctgcc
      361 gggcggcaat ctgggtgccc agaatctgag cgaaagcgcc gccgtgaagg agatcctgaa
      421 agagcaggag aaccgcaaag gcctgattgc cgccatttgt gcaggcccga ccgcactgct
      481 ggcccatgaa attggcttcg gtagcaaggt taccacccac ccgctggcca aagataagat
      541 gatgaacggc ggccactaca cctacagcga aaaccgcgtg gaaaaagatg gcctgattct
      601 gaccagccgc ggtccgggta ccagctttga attcgccctg gcaatcgttg aggccctgaa
      661 cggcaaagaa gtggccgccc aggtgaaagc cccgctggtt ctgaaagact aataaggatc
      721 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      781 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
      841 ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct
      901 ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga
      961 acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg
     1021 gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     1081 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     1141 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     1201 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     1261 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1441 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1501 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1561 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1621 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1681 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1741 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1801 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1861 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1921 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1981 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2041 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2101 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2161 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2221 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2281 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2341 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2581 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2701 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2761 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2821 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2881 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2941 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3001 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3061 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3241 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     3301 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3361 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3541 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3601 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3661 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     3721 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     3781 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     3841 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     3901 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     3961 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     4021 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4081 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4141 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     4201 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     4261 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     4321 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     4381 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     4441 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     4501 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     4561 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     4621 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4681 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4741 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4801 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4861 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4921 tctcaagggc atcggtcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4981 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5041 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5101 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5161 aaccgcacct gtgg
//
