LOCUS       sequence-243310-        7602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse TRAF3IP3 ∆TM in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  TITLE     Sun Lab TRAF3ip and Tbkbp1 expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7602)
  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            "Unpublished"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: pcDNA-mTRAF3ip3∆TM-CFP
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     692..711
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        692..710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2249..2251,2252..2254,2255..2968)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2294..2315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2555..2574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2902..2923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2984..3002)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2985..3002)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3022..3039)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3028..3252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3298..3726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3385..3404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3595..3616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3735..3755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3740..4069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3982..4001
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4136..4930
                     /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(4190..4209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4800..4819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5104..5225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5141..5160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5195..5214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5274..5290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5274..5290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5287..5309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5298..5314
                     /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(5322..5328,5329..5346,5347..5352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5367..5388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5505..5522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5676..6264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5756..5775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6435..7226,7227..7295))
                     /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     7058..7077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7296..7400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7475..7494)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
       61 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      661 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      721 cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctggaat tcaccatgat
      781 cagctcagac tcgaggtcct cccctggctt ggctcggtgg gctgagagct atgaggccaa
      841 gtgtgagcga cgtcaggaga cccgagaaaa ccgtcgccgc cgccgcaatg agaccacttg
      901 ccgccagccg gggaaggtgc tgaggaccca acacaaggaa cggcttcagg gagctcggca
      961 gctacagttc ctcaaaagga gaaacctgga agaagagaag aaaggccaag ccagggaaca
     1021 agggccctcc agcaggacag atggaggaac aggccaagtg tctatcctca aggagtcctt
     1081 gcctggtgcc aacaaggcct ctttccctgg acagcaggaa acagggataa gctctgaggt
     1141 tttcccggcc ttacatcatt cctcctcagg catccagagg gatctgggtg gccaccatgc
     1201 ctcccatggg agagcctttc caccacagga ctctgacatc aagaagccac acagacagca
     1261 ccggggcact cagacaaagg cagaagaggc attgccaaca ataaagaatg atgccagtca
     1321 gcaaaccaaa tgtggagttg ctgtcctaga caaggacatc attcaactct cggaatacct
     1381 caaagaagcc ttacataggg agctgatcct aaaaaagaaa atggtgattc tccaggacct
     1441 cctgcctgcc ctgatccggg cctctgacag ctcctggaag ggacagctta atgaagacaa
     1501 gctgaagggc aaactaagat ccctagaaaa ccagctctac acctgcctgc agaaacactc
     1561 cccctgggga atgaaaaagg tgctgctgga gatggaagac cagagaagca gctatgagca
     1621 gaaggctaag gcgtcactgc agaaagtgct ggaggagaaa atgtgtgcag agcagcagct
     1681 acagagggca cagctgtccc tggctttggc agagcagaag tgtcaggagt ggaagagtca
     1741 gtacgaggct ctgaaggagg actggaggac cctaggggac cagcacagag agctggagag
     1801 ccaactgcat gtgcttcagt ccaaactaca gggagcagac agcagagacc cacagatgag
     1861 ccaggccctg cagcttttag aaaatgagca ccaggagctg cagaccaaac tcgagagcct
     1921 gcagggagac ggagagcagc agagctctga gacccaggac ctacaagatc aactaaagaa
     1981 gtcagaggaa gagaaacagg ccctggtgag caaagtacaa cagctacaga gtctgcttca
     2041 gaatcaatcc ttacaacttc aagaacagga gaagctctta aagaaagatc agggtctgcc
     2101 tgtgtggaat ccaaagctct cccttgatga agtgaagcct gagggcacaa gaaaggagaa
     2161 agaggaggag ctcagagacc aactacaaaa ggagacattt cagctccaag tcaaggaaaa
     2221 ggagctgcag tgtggtcaat gggtcgagat ggtgagcaag ggcgaggagc tgttcaccgg
     2281 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc
     2341 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2401 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg
     2461 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2521 aggctacgtc caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2581 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2641 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt
     2701 ctatatcacc gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa
     2761 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2821 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2881 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2941 tctcggcatg gacgagctgt acaagtaatc tagagggccc tattctatag tgtcacctaa
     3001 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3061 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3121 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3181 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3241 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3301 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3361 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3421 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3481 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3541 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3601 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3661 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3721 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4081 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4141 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4201 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4261 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4321 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4381 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4441 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4501 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4561 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4621 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4681 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4741 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4801 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4861 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4921 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4981 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5041 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5101 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5221 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5281 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5341 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5401 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5461 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6541 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7021 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7081 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7141 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7201 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7321 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7381 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7441 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7501 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7561 tttaagctac aacaaggcaa ggcttgaccg acaattgcat ga
//
