LOCUS       sequence-489492-        9056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 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 9056)
  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 9056)
  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
FEATURES             Location/Qualifiers
     source          1..9056
                     /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             5012..5722
                     /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(5157..5175)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5425..5444
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5844..5965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5881..5900)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5935..5954
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5972..6427)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6109..6130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6321..6340
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6454..6558
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6560..6917
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6560..6751
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6583..6603)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6768..6903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6952..7746
                     /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             6952..7746
                     /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(7006..7025)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7616..7635
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7934..7953)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7978..8025
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8354..8942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8843..8862
                     /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 gcagcgacag
     4021 cttcgtgatg ctgctggaga gcgaaagcgg tccagagagc gtgccaccag gtagcctgag
     4081 caacgtgagc gacaacgctg gtgtgcaggg tagcccgctg gtgaacaact acggtcaggg
     4141 tagcccggct gctaacagca gcgtgagccc aagcggtgaa gatctgattt ttcacccggt
     4201 gagcgtgctg gtgctgaagg tgaacgaggt gagcttcggc atcgaagtgc gtggcgagga
     4261 cctgaccgtg gctctgcagg ccgaggaact gaccctgcag cagctgggca ccgtgggtct
     4321 gtggcagttt ctgcacggcc agtgcccggg tacctgcttc caggaaagca gcaccctgaa
     4381 aaccggtcac atccgtccag ctgtgggtct gcgttttgaa gtgggtccag gtgctgctgt
     4441 gcacagccca ctggctagcc agaacggctt cctgcacctg ctgctgcacg gttgcgatct
     4501 ggagctgctg accagcgtgc tgagcggcct gggtccgttt ctggaagacg aggaaattcc
     4561 ggtggtggtg ccgatgcaga ttgagctgct gaacagcagc atcaccctga aggacgatat
     4621 tccaccaatc tatccgacca gcccaggtcc aattccaatc accctggcca tggagcacgt
     4681 ggtgctgaaa cgcagcgacg atggcgtgtt ccacatcggt gctgctgctc aggacaagcc
     4741 aagcgcggaa gtgctgaaga gcgagaaacg tcagccgccg aaagaacagg tgtttctggt
     4801 gccgaccggt gaagtgttcg agcagcaggt gaaggagctg ccgattctgc agaaggaact
     4861 gatcgaaacc aaacaggctc tggccaacgc gaaccaggac aaggaaaaac tgctgcaaga
     4921 gattcgtaag tataacccat tctttgaact gggtggatct ggaggtcgaa ttctgcagtc
     4981 gacggtaccg cgggcccggg atccaccggt catggtgagc aagggcgagg aggataacat
     5041 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     5101 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     5161 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     5221 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     5281 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     5341 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     5401 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     5461 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     5521 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     5581 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     5641 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     5701 catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc cataccacat
     5761 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5821 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5881 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5941 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     6001 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6061 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6121 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6181 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6241 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6301 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6361 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6421 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6541 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6661 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6721 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6781 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6841 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6901 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     6961 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     7021 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     7081 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     7141 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     7201 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     7261 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7321 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7381 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7441 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7501 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7561 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7621 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7681 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7741 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     7801 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7861 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     7921 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     7981 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     8041 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     8101 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     8161 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     8221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9001 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
