LOCUS       sequence-379803-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Presenilin Associated Rhomboid Like (PARL) D319A 
            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  JOURNAL   unpublished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            "unpublished"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-D319A-FLAG C-t
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1999..2018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2403..2454
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     2405..2425
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2499..2518
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2499..2517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3711..3734
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3753..3771)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3754..3771)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3791..3808)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3797..4021
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4067..4495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4154..4173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4364..4385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4504..4524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4509..4838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4689..4824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4751..4770
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4905..5699
                     /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(4959..4978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5569..5588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5873..5994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5910..5929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5964..5983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6043..6059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6043..6059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6056..6078)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6067..6083
                     /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(6091..6097,6098..6115,6116..6121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6136..6157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     1981 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     2041 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     2101 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     2161 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     2221 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2281 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2341 gtatgttccc atagtaacgc caatagggac tttccaaaat gtcgtaacaa ctccgcccca
     2401 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2461 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2521 acccaagatt ggtacccggg aattcgcggc cgcgtcgacc agaggttggg aagatggcgt
     2581 ggcgaggctg ggcgcagaga ggctggggct gcggccaggc gtggggtgcg tcggtgggcg
     2641 gccgcagctg cgaggagctc actgcggtcc taaccccgcc gcagctcctc ggacgcaggt
     2701 ttaacttctt tattcaacaa aaatgcggat tcagaaaagc acccaggaag gttgaacctc
     2761 gaagatcaga cccagggaca agtggtgaag catacaagag aagtgctttg attcctcctg
     2821 tggaagaaac agtcttttat ccttctccct atcctataag gagtctcata aaacctttat
     2881 tttttactgt tgggtttaca ggctgtgcat ttggatcagc tgctatttgg caatatgaat
     2941 cactgaaatc cagggtccag agttattttg atggtataaa agctgattgg ttggatagca
     3001 taagaccaca aaaagaagga gacttcagaa aggagattaa caagtggtgg aataacctaa
     3061 gtgatggcca gcggactgtg acaggtatta tagctgcaaa tgtccttgta ttctgtttat
     3121 ggagagtacc ttctctgcag cggacaatga tcagatattt cacatcgaat ccagcctcaa
     3181 aggtcctttg ttctccaatg ttgctgtcaa cattcagtca cttctcctta tttcacatgg
     3241 cagcaaatat gtatgttttg tggagcttct cttccagcat agtgaacatt ctgggtcaag
     3301 agcagttcat ggcagtgtac ctatctgcag gtgttatttc caattttgtc agttacctgg
     3361 gtaaagttgc cacaggaaga tatggaccat cacttggtgc atctggtgcc atcatgacag
     3421 tcctcgcagc tgtctgcact aagatcccag aagggaggct tgccattatt ttccttccga
     3481 tgttcacgtt cacagcaggg aatgccctga aagccattat cgccatggca acagcaggaa
     3541 tgatcctggg atggaaattt tttgatcatg cggcacatct tgggggagct ctttttggaa
     3601 tatggtatgt tacttacggt catgaactga tttggaagaa cagggagccg ctagtgaaaa
     3661 tctggcatga aataaggact aatggcccca aaaaaggagg tggctctaag gactacaagg
     3721 acgacgatga caagtgatct agagggccct attctatagt gtcacctaaa tgctagagct
     3781 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3841 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3901 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3961 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4021 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4081 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4141 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4201 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4261 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4321 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4381 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4441 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4501 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4621 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4681 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4741 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4801 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4861 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4921 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4981 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5041 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5101 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5161 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5221 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5281 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5341 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5401 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5461 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5521 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5581 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5641 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5701 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5761 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5821 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5881 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5941 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6001 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6061 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6121 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6181 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6241 aggcg
//
