LOCUS       sequence-190815-        7659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Psoriasis-Linked mutant CARD14/CARMA2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  AUTHORS   Scudiero I, Mazzone P, D'Andrea LE, Ferravante A, Zotti T, Telesio 
            G, De Rubis G, Reale C, Pizzulo M, Muralitharan S, Vito P, Stilo R
  TITLE     CARMA2sh and ULK2 control pathogen-associated molecular patterns 
            recognition in human keratinocytes: psoriasis-linked CARMA2sh 
            mutants escape ULK2 censorship.
  JOURNAL   Cell Death Dis. 2017 Feb 23;8(2):e2627. doi: 10.1038/cddis.2017.51.
  PUBMED    28230860
REFERENCE   2  (bases 1 to 7659)
  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: "Cell Death 
            Dis."; date: "2017-02-23"; volume: "8(2):e2627. doi"; pages: " 
            10.1038/cddis.2017.51"
COMMENT     Sequence Label: pcDNA3-CARMA2sh-E138A
FEATURES             Location/Qualifiers
     source          1..7659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             692..718
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3016..3033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3022..3246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3292..3720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3589..3610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3729..3749)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3734..4063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3914..4049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3976..3995
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4130..4924
                     /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(4184..4203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4794..4813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5098..5219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5135..5154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5189..5208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5268..5284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5281..5303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5292..5308
                     /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(5316..5322,5323..5340,5341..5346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5499..5516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6429..7220,7221..7289))
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7469..7488)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttaccatggg atacccatac gatgttccag attacgctgt
      721 ggtaccagga tccagtgtgg tggaattcat gggggaactg tgccgcaggg actccgcact
      781 cacggcactg gacgaggaga cactgtggga gatgatggag agccaccgcc acaggatcgt
      841 acgctgcatc tgccccagcc gcctcacccc ctacctgcgc caggccaagg tgctgtgcca
      901 gctggacgag gaggaggtgc tgcacagccc ccggctcacc aacagcgcca tgcgggccgg
      961 gcacttgctg gatttgctga agactcgagg gaagaacggg gccatcgcct tcctggagag
     1021 cctgaagttc cacaaccctg acgtctacac cctggtcacc gggctgcagc ctgatgttga
     1081 cttcagtaac tttagcggtc tcatggagac atccaagctg accgagtgcc tggctggggc
     1141 catcggcagc ctgcaggagg cgctgaacca ggaaaagggg cagaaggagg tgctgctgcg
     1201 gcggtgccag cagctgcagg agcacctggg cctggccgag acccgtgccg agggcctgca
     1261 ccagctggag gctgaccaca gccgcatgaa gcgtgaggtt agcgcacact tccatgaggt
     1321 gctgaggctg aaggacgaga tgctcagcct ctcgctgcac tatagcaatg cgctgcagga
     1381 aaaggagctg gccgcctcac gctgccgcag cctgcaggag gagctgtatc tactgaagca
     1441 ggagctgcag cgagccaaca tggtttcctc ctgtgagctg gaattgcaag agcagtccct
     1501 gaggacagcc agcgaccagg agtccgggga tgaggagctg aaccgcctga aggaggagaa
     1561 tgagaaactg cgctcgctga ctttcagcct ggcggagaag gacattctgg agcagagcct
     1621 ggacgaggcg cgggggagcc gacaggagct ggtggagcgc atccactcgc tgcgggagcg
     1681 ggccgtggct gccgagaggc agcgagagca gtactgggaa gagaaggaac agaccctgct
     1741 gcagttccag aagagtaaga tggcctgcca actctacagg gagaaggtga atgcgctgca
     1801 ggcccaggtg tgcgagctgc agaaggagcg agaccaggcg tactccgcga gggacagtgc
     1861 tcagagggag atttcccaga gcctggtgga gaaggactcc ctccgcaggc aggtgttcga
     1921 gctgacggac caggtctgcg agctgcgcac acagcttcgc cagctgcagg cagagcctcc
     1981 gggtgtgctc aagcaggaag ccaggaccag ggagccctgt ccacgggaga agcagcggct
     2041 ggtgcggatg catgccatct gccccagaga cgacagcgac tgcagcctcg tcagctccac
     2101 agagtctcag ctcttgtcgg acctgagtgc cacgtccagc cgcgagctgg tggacagctt
     2161 ccgctccagc agccccgcgc cccccagcca gcagtccctg tacaagcggg tggccgagga
     2221 cttcggggaa gaaccctggt ctttcagcag ctgcctggag atcccggagg gagacccggg
     2281 agccctgccg ggagctaagg caggcgaccc acacctggat tatgagctcc tagacacggc
     2341 agaccttccg cagctggaaa gcagcctgca gccagtctcc cctggaaggc ttgatgtctc
     2401 ggagagcggc gtcctcatgc ggcggaggcc agcccgcagg atcctgagcc aggtcaccat
     2461 gctggcgttc cagggggatg cattgctgga gcagatcagc gtcatcggcg ggaacctcac
     2521 gggcatcttc atccaccggg tcaccccggg ctcggcggcg gaccagatgg ccttgcgccc
     2581 gggcacccag attgtgatgg ttgattacga agcctcagag cccttgttca aggcagtcct
     2641 ggaggacacg accctggagg aggccgtggg gcttctcagg agggtggacg gcttctgctg
     2701 cctgtctgtg aaggtcaaca cggacggtta taagaggcta ctccaggacc tggaggccaa
     2761 agtggcgacc tcgggggact cattctacat ccgggtcaac ctggccatgg agggcagggc
     2821 caaaggggag ctgcaggtgc attgcaacga ggtcctgcac gtcaccgaca ccatgttcca
     2881 gggctgcggc tgctggcatg cccaccgcgt gaactcttac accatgaagg atactgccgc
     2941 gcacggcacc atccccaact actccaggtg agcggccgct cgagtctaga gggcccgttt
     3001 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3061 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3121 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3181 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3241 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3601 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3661 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3721 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3901 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3961 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4021 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4081 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4141 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4201 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4261 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4321 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4381 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4441 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4501 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4561 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4621 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4681 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4741 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4801 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4861 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4921 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4981 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5041 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5221 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7441 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7501 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7561 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     7621 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
