LOCUS       sequence-379806-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Presenilin Associated Rhomboid Like (PARL) D319N 
            mutant .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6245)
  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            "unpublished"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: pcDNA3-PARL-D319N-FLAG C-t
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(93..110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(264..852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(344..363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1023..1814,1815..1883))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1646..1665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1884..1988)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2063..2082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2467..2518
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     2469..2489
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2563..2582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2563..2581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3775..3798
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3817..3835)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3818..3835)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3855..3872)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3861..4085
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4131..4559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4428..4449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4568..4588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4573..4902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4753..4888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4815..4834
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4969..5763
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5023..5042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5633..5652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5937..6058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5974..5993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6028..6047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6107..6123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6107..6123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6120..6142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6131..6147
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6155..6161,6162..6179,6180..6185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6200..6221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
       61 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      181 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      241 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      301 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      421 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      601 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      661 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      721 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      781 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      841 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      901 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      961 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1021 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1081 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1141 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1201 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1261 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1321 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1381 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1441 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1501 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1561 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1621 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1681 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1741 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1801 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1861 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1921 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1981 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     2041 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     2101 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     2161 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     2221 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     2281 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2341 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2401 tgacgtatgt tcccatagta acgccaatag ggactttcca aaatgtcgta acaactccgc
     2461 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     2521 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     2581 ggagacccaa gattggtacc cgggaattcg cggccgcgtc gaccagaggt tgggaagatg
     2641 gcgtggcgag gctgggcgca gagaggctgg ggctgcggcc aggcgtgggg tgcgtcggtg
     2701 ggcggccgca gctgcgagga gctcactgcg gtcctaaccc cgccgcagct cctcggacgc
     2761 aggtttaact tctttattca acaaaaatgc ggattcagaa aagcacccag gaaggttgaa
     2821 cctcgaagat cagacccagg gacaagtggt gaagcataca agagaagtgc tttgattcct
     2881 cctgtggaag aaacagtctt ttatccttct ccctatccta taaggagtct cataaaacct
     2941 ttatttttta ctgttgggtt tacaggctgt gcatttggat cagctgctat ttggcaatat
     3001 gaatcactga aatccagggt ccagagttat tttgatggta taaaagctga ttggttggat
     3061 agcataagac cacaaaaaga aggagacttc agaaaggaga ttaacaagtg gtggaataac
     3121 ctaagtgatg gccagcggac tgtgacaggt attatagctg caaatgtcct tgtattctgt
     3181 ttatggagag taccttctct gcagcggaca atgatcagat atttcacatc gaatccagcc
     3241 tcaaaggtcc tttgttctcc aatgttgctg tcaacattca gtcacttctc cttatttcac
     3301 atggcagcaa atatgtatgt tttgtggagc ttctcttcca gcatagtgaa cattctgggt
     3361 caagagcagt tcatggcagt gtacctatct gcaggtgtta tttccaattt tgtcagttac
     3421 ctgggtaaag ttgccacagg aagatatgga ccatcacttg gtgcatctgg tgccatcatg
     3481 acagtcctcg cagctgtctg cactaagatc ccagaaggga ggcttgccat tattttcctt
     3541 ccgatgttca cgttcacagc agggaatgcc ctgaaagcca ttatcgccat gaacacagca
     3601 ggaatgatcc tgggatggaa attttttgat catgcggcac atcttggggg agctcttttt
     3661 ggaatatggt atgttactta cggtcatgaa ctgatttgga agaacaggga gccgctagtg
     3721 aaaatctggc atgaaataag gactaatggc cccaaaaaag gaggtggctc taaggactac
     3781 aaggacgacg atgacaagtg atctagaggg ccctattcta tagtgtcacc taaatgctag
     3841 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3901 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3961 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4021 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4081 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4141 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4201 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4261 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4321 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4381 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4441 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4501 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4561 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4621 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4741 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4801 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4861 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4921 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4981 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5041 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5101 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5161 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5221 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5281 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5341 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5401 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5461 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5521 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5581 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5641 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5701 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5761 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6061 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6241 ccgct
//
