LOCUS       sequence-492341-        9038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 
            lacking its LIR motif as a C-terminal fusion to RFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9038)
  AUTHORS   Hanna MG, Rodriguez Cruz HO, Fujise K, Wu Y, Xu CS, Pang S, Li Z, 
            Monetti M, De Camilli P
  TITLE     BLTP3A is associated with membranes of the late endocytic pathway 
            and is an effector of CASM.
  JOURNAL   EMBO J (2025) 44: 6168 - 6195
  PUBMED    39386594
REFERENCE   2  (bases 1 to 9038)
  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: "EMBO J 
            (2025) 44: 6168 - 6195"
COMMENT     Sequence Label: pmCh-N1 BLTP3A LIR mutant
FEATURES             Location/Qualifiers
     source          1..9038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4994..5704
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5139..5157)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5407..5426
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5826..5947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5863..5882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5917..5936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5954..6409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6091..6112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6303..6322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6436..6540
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6542..6899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6542..6733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6565..6585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6750..6885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6934..7728
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6934..7728
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6988..7007)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7598..7617
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7916..7935)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7960..8007
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8336..8924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8825..8844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagac cggtcgccac catggcgggt atcatcaaaa aacaaatcct
      661 gaaacacctg agccgcttta ccaagaacct gagcccagac aaaatcaacc tgagcaccct
      721 gaaaggcgag ggtcagctga ccaacctgga gctggacgag gaagtgctgc agaacgtgct
      781 ggagctgccg acctggctgg ccattacccg cgtgtactgc aaccgtgcga gcatccgcat
      841 tcagtggacc aagctgaaaa cccacccgat ctgcctgtgc ctggataagg tggaggtgga
      901 aatgaaaacc tgcgaagacc cgcgtccgcc aaacggtcag agcccaattg cgctggctag
      961 cggtcagagc gagtacggtt ttgccgaaaa ggtggtggag ggcatgttca tcattgtgaa
     1021 cagcatcacc attaagatcc acagcaaagc tttccacgcc agctttgaac tgtggcagct
     1081 gcagggctat agcgtgaacc cgaactggca gcagagcgat ctgcgtctga cccgtatcac
     1141 cgacccgtgc cgtggtgaag tgctgacctt taaggagatt acctggcaga ccctgcgtat
     1201 cgaggccgac gcgaccgata acggcgacca ggacccggtt accaccccac tgcgcctgat
     1261 taccaaccag ggtcgtattc agatcgccct gaagcgtcgc accaaagatt gcaacgtgat
     1321 cagcagcaaa ctgatgttcc tgctggacga tctgctgtgg gtgctgaccg acagccagct
     1381 gaaggccatg atgaaatacg cggaaagcct gagcgaggct atggaaaaga gcgcccacca
     1441 gcgtaaaagc ctggctccag agccggtgca gatcacccca ccagctccaa gcgcccagca
     1501 gagctgggct caggccttcg gcggtagcca gggcaacagc aacagcagca gcagccgcct
     1561 gagccagtac ttcgaaaagt ttgatgtgaa agagagcagc tatcacctgc tgattagccg
     1621 tctggacctg cacatctgcg acgatagcca gagccgtgag ccaggtgtga gcgccaaccg
     1681 tctgatgggc ggtgcgatgc agctgacctt ccgtaagatg gcttttgatt actatccgtt
     1741 ccactgggcc ggcgacagct gcaaacactg ggtgcgccac tgcgaggcta tggaaacccg
     1801 tggccagtgg gcccagaagc tggtgatgga atttcagagc aagatggaga aatggcacga
     1861 ggaaaccggc ctgaaaccac catggcacct gggtgtggat agcctgttcc gtcgtaaggc
     1921 tgacagcctg agcagcccgc gtaaaaaccc actggagcgc agcccgagcc agggccgtca
     1981 gccagctttc cagccaccag cttggaaccg tctgcgcagc agctgcatgg tggtgcgcgt
     2041 ggacgatctg gatattcacc aggtgagcac cgctggtcag ccgagcaaga aaccgagcac
     2101 cctgctgagc tgcagccgta agctgcacaa cctgccgacc caggtgagcg cgattcacat
     2161 cgagtttacc gaatactatt tcccggacaa ccaggaactg ccagtgccgt gcccgaacct
     2221 gtacatccag ctgaacggcc tgacctttac catggacccg gtgagcctgc tgtggggtaa
     2281 cctgttttgc ctggacctgt accgtagcct ggagcagttc aaggccattt ataaactgga
     2341 agatagcagc cagaaagacg agcacctgga tatccgcctg gacgcttttt ggctgaaggt
     2401 gagcttcccg ctggagaaac gtgaacgtgc tgagctgcac cgtccacagg ccctggtgtt
     2461 cagcgcgagc ggtatgatcg ctaccaacac ccgtcacgct ccacactgca gctgcagcga
     2521 tctgcagagc ctgttccgtg gttttgccgc ggctgagttc tttcacagca actatgacca
     2581 ctttccgaag gtgccgggcg gtttcagcct gctgcacatg ctgtttctgc accacgcctt
     2641 ccagatggac agctgcctgc cacagccaaa caccctgcca ccacagcgtc cgaaggcgag
     2701 ctgggatctg tggagcgtgc actttaccca gattagcctg gactttgaag gcaccgagaa
     2761 cttcaaaggt cacaccctga acttcgtggc gccgtttccg ctgagcatct gggcttgcct
     2821 gccactgcgt tggcagcagg ctcaggctcg taagctgctg ctggctagcg agggccgcct
     2881 gaaaccaagc gccagcttcg gtagcccggt gcagagcgaa gccctggctc cagatagcat
     2941 gagccaccca cgtagcaaga ccgagcacga cctgaaaagc ctgagcggcc tgaccgaagt
     3001 gatggagatc ctgaaggagg gcagcagcgg tatggacaac aaaggtccgc tgaccgagct
     3061 ggaggacgtg gccgatgtgc acatgctggt gcacagccca gctcacgtgc gtgtgcgcct
     3121 ggatcactac cagtatctgg ccctgctgcg cctgaaggaa gtgctgcagc gtctgcagga
     3181 gcagctgacc aaagacaccg aaagcatgac cggcagcccg ctgcagaacc agaccgcgtg
     3241 cattggtgtg ctgttcccaa gcgccgaggt ggcgctgctg atgcacccag ctccaggtgc
     3301 tgtggacgct gatagcgccg gtagcgatag caccagcctg gtggacagcg aactgagccc
     3361 gagcgaagac cgcgagctga agagcgatgc tagcagcgac cagggtccag ctagcccaga
     3421 gaaagtgctg gaggaaagca gcatcgagaa ccaggatgtg agccaggaac gtccgcacag
     3481 caacggcgag ctgcaggaca gcggtccact ggcgcagcag ctggctggca agggtcacga
     3541 agcggtggag agcctgcagg ctaagaaact gagccgcacc caggccagca gcagcccagc
     3601 cgcgctgaag ccaccagctg gtcgtgaaac cgcggtgaac ggccagggcg agctgattcc
     3661 gctgaaaaac atcgagggtg aactgagcag cgcgattcac atgaccaagg atgctaccaa
     3721 agaagccctg cacgcgacca tggacctgac caaggaggcc gtgagcctga ccaaagatgc
     3781 gttcagcctg ggccgtgacc gcatgaccag caccatgcac aagatgctga gcctgccgcc
     3841 ggctaaggaa ccgatggcca aaaccgatga gggtgtggct gctccggtga gcggtggtgc
     3901 tgctcgtctg cgcttcttta gcatgaaacg caccgtgagc cagcagagct ttgatggcgt
     3961 gagcctggac agcagcggtc cggaagatcg tatcagcgtg gacagcgatg gcagcctgga
     4021 gagcgaaagc ggtccagaga gcgtgccacc aggtagcctg agcaacgtga gcgacaacgc
     4081 tggtgtgcag ggtagcccgc tggtgaacaa ctacggtcag ggtagcccgg ctgctaacag
     4141 cagcgtgagc ccaagcggtg aagatctgat ttttcacccg gtgagcgtgc tggtgctgaa
     4201 ggtgaacgag gtgagcttcg gcatcgaagt gcgtggcgag gacctgaccg tggctctgca
     4261 ggccgaggaa ctgaccctgc agcagctggg caccgtgggt ctgtggcagt ttctgcacgg
     4321 ccagtgcccg ggtacctgct tccaggaaag cagcaccctg aaaaccggtc acatccgtcc
     4381 agctgtgggt ctgcgttttg aagtgggtcc aggtgctgct gtgcacagcc cactggctag
     4441 ccagaacggc ttcctgcacc tgctgctgca cggttgcgat ctggagctgc tgaccagcgt
     4501 gctgagcggc ctgggtccgt ttctggaaga cgaggaaatt ccggtggtgg tgccgatgca
     4561 gattgagctg ctgaacagca gcatcaccct gaaggacgat attccaccaa tctatccgac
     4621 cagcccaggt ccaattccaa tcaccctggc catggagcac gtggtgctga aacgcagcga
     4681 cgatggcgtg ttccacatcg gtgctgctgc tcaggacaag ccaagcgcgg aagtgctgaa
     4741 gagcgagaaa cgtcagccgc cgaaagaaca ggtgtttctg gtgccgaccg gtgaagtgtt
     4801 cgagcagcag gtgaaggagc tgccgattct gcagaaggaa ctgatcgaaa ccaaacaggc
     4861 tctggccaac gcgaaccagg acaaggaaaa actgctgcaa gagattcgta agtataaccc
     4921 attctttgaa ctgggtggat ctggaggtcg aattctgcag tcgacggtac cgcgggcccg
     4981 ggatccaccg gtcatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     5041 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     5101 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5161 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     5221 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     5281 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5341 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     5401 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     5461 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     5521 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     5581 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     5641 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     5701 gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     5761 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5821 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5941 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6361 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     6421 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6481 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6541 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6661 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6721 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     6901 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6961 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     7021 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     7081 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     7141 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     7201 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     7261 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     7321 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     7381 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     7441 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     7501 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7561 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7621 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7681 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7741 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     7801 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     7861 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     7921 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7981 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     8041 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     8101 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     8161 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     8221 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8281 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8341 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8401 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8461 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8521 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8581 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8641 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8701 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8761 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8821 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8881 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8941 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9001 cccctgattc tgtggataac cgtattaccg ccatgcat
//
