LOCUS       sequence-489502-        8828 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 1-1364
            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 8828)
  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 8828)
  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 1-1364
FEATURES             Location/Qualifiers
     source          1..8828
                     /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             4784..5494
                     /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(4929..4947)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5197..5216
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5616..5737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5653..5672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5707..5726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5744..6199)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5881..5902)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6093..6112
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6226..6330
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6332..6689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6332..6523
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6355..6375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6540..6675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6724..7518
                     /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             6724..7518
                     /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(6778..6797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7388..7407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7706..7725)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7750..7797
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8126..8714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8615..8634
                     /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 gctggtggat ctggaggtcg
     4741 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcatggtga gcaagggcga
     4801 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     4861 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     4921 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     4981 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     5041 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     5101 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     5161 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     5221 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     5281 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     5341 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     5401 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     5461 ctccaccggc ggcatggacg agctgtacaa gtaagcggcc gcgactctag atcataatca
     5521 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     5581 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     5641 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5701 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     5761 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5821 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5881 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5941 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6001 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6061 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6121 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6181 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6301 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     6361 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6421 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6481 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6541 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6601 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6661 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     6721 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6781 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6841 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6901 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6961 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     7021 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     7081 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     7141 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     7201 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     7261 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     7321 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     7381 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7441 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7501 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     7561 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     7621 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     7681 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     7741 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     7801 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     7861 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     7921 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     7981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8821 ccatgcat
//
