LOCUS       sequence-489604-        8717 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 1-1327
            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 8717)
  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   bioRxiv [Preprint]. 2025 Apr 3:2024.09.28.615015. doi: 
            10.1101/2024.09.28.615015.
  PUBMED    39386594
REFERENCE   2  (bases 1 to 8717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Apr 3:2024.09.28.615015. doi: 
            10.1101/2024.09.28.615015."
COMMENT     Sequence Label: pmCh-N1 BLTP3A 1-1327
FEATURES             Location/Qualifiers
     source          1..8717
                     /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             4673..5383
                     /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(4818..4836)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5086..5105
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5505..5626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5542..5561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5596..5615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5633..6088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5770..5791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5982..6001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6115..6219
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6221..6578
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6221..6412
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6244..6264)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6429..6564
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6613..7407
                     /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             6613..7407
                     /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(6667..6686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7277..7296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7595..7614)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7639..7686
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8015..8603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8504..8523
                     /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 agggtggatc
     4621 tggaggtcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcatggtgag
     4681 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     4741 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4801 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4861 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4921 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4981 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     5041 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     5101 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     5161 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     5221 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     5281 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     5341 gggccgccac tccaccggcg gcatggacga gctgtacaag taagcggccg cgactctaga
     5401 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5461 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     5641 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5701 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5761 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5821 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5881 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5941 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6001 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6061 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6181 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     6241 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6301 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6421 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6481 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6541 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     6601 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6661 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6721 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6781 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6841 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6901 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6961 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     7021 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7081 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7141 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7201 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7261 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7321 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7381 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     7441 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7501 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7561 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     7621 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     7681 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     7741 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     7801 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     7861 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7921 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7981 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8041 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8101 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8161 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8221 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8281 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8341 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8401 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8461 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8521 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8581 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8641 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8701 gtattaccgc catgcat
//
