LOCUS       sequence-489591-        9128 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 
            chimera-1 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 9128)
  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 9128)
  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 BLTP3 chimera-1
FEATURES             Location/Qualifiers
     source          1..9128
                     /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             5084..5794
                     /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(5229..5247)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5497..5516
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5916..6037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5953..5972)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6007..6026
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6044..6499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6181..6202)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6393..6412
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6526..6630
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6632..6989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6632..6823
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6655..6675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6840..6975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7024..7818
                     /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             7024..7818
                     /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(7078..7097)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7688..7707
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8006..8025)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8050..8097
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8426..9014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8915..8934
                     /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 ggcatgagcg tgagcgtgaa
     1021 cagcatcgtg attcgtatcg gtgccaaggc gtttaacgcc agcttcgaac tgagccagct
     1081 gcgcatctac agcgtgaacg cgcactggga gcacggcgat ctgcgtttca cccgcatcca
     1141 ggacccgcag cgtggtgagg tgctgacctt taaggaaatt aactggcaga tgattcgcat
     1201 cgaagctgac gccacccaga gcagccacct ggagatcatg tgcgctccgg tgcgtctgat
     1261 caccaaccag agcaaaattc gcgtgaccct gaagcgtcgc ctgaaagatt gcaacgtgat
     1321 cgccaccaag ctggtgctga ttctggacga tctgctgtgg gtgctgaccg acagccagct
     1381 gaaggctatg gtgcagtacg ccaaaagcct gagcgaggct atcgaaaaga gcaccgaaca
     1441 gcgtaaaagc atggccccag agccaaccca gagcagcacc gtggtggcta gcgcccagca
     1501 ggtgaagacc acccagacca gcaacgcgcc ggacgtgaac gatgctattg tgaaactgtt
     1561 taacgacttc gatgtgaagg aaaccagcca ccacctggtt atcagccacc tggatctgca
     1621 catctgcgac gatattcacg cgaaggagaa agaaagcaac cgtcgcatca ccggcggtgc
     1681 tatgcagctg agcttcaccc agctgaccat tgattactat ccgtatcaca aagcgggcga
     1741 cagctgcaac cactggatgt acttcagcga cgcgaccaag accaaaaacg gttgggctaa
     1801 cgaactgctg cacgagtttg aatgcaacgt ggagatgctg aagcaggcgg tgaaagatca
     1861 caacgtgggc agcccgccga agagcccaac ccacgctagc ccacagcaca cccagaccga
     1921 gaaggactat ccgctgaaag gtacctgccg taccccgagc gtgctgagcc agcagagcaa
     1981 ggccaaactg atgagcagca gcgtggtggt gcgtctggcg gacttcaaca tctaccaggt
     2041 gagcaccgct gaacagtgcc gcagcagccc gaaaagcatg atctgctgca acaagaaaag
     2101 cctgtatctg ccgcaggaga tgagcgcggt gtacatcgag ttcaccgaat actattaccc
     2161 ggacggcaag gattttccga ttccgagccc gaacctgtat agccagctga acgctctgca
     2221 gttcaccgtg gatgaacgta gcatcctgtg gctgaaccag ttcctgctgg acctgaagca
     2281 gagcctgaac cagtttatgg ccgtgtacaa gctgaacgat aacagcaaaa gcgacgagca
     2341 cgtggatgtg cgtgtggacg gtctgatgct gaaatttgtg attccgagcg aggtgaagag
     2401 cgaatgccac caggaccagc cacgtgccat tagcatccag agcagcgaaa tgatcgcgac
     2461 caacacccgt cactgcccaa actgccgtca cagcgatctg gaggcgctgt ttcaggactt
     2521 caaggactgc gatttcttta gcaaaaccta taccagcttt ccgaagagct gcgataactt
     2581 caacctgctg cacccgatct ttcagcgtca cgcccacgag caggacacca agatgcacga
     2641 aatctacaaa ggcaacatta ccccgcagct gaacaagaac accctgaaaa ccagcgccgc
     2701 taccgacgtg tgggccgtgt acttcagcca gttctggatc gactacgagg gtatgaagag
     2761 cggcaaaggt cgtccgatta gctttgtgga tagcttcccg ctgagcattt ggatctgcca
     2821 gccgacccgc tatgccgaga gccagaagga accgcagacc tgcaaccagg tgagcctgaa
     2881 caccagccag agcgaaagca gcgacctggc tggccgtctg aagcgcaaga aactgctgaa
     2941 agagtattac agcaccgaga gcgaaccgct gaccaacggc ggtcagaaac cgagcagcag
     3001 cgataccttc tttcgtttca gcccgagcag cagcgaagcc gacatccacc tgctggtgca
     3061 cgtgcacaag cacgtgagca tgcagattaa ccactatcag tacctgctgc tgctgtttct
     3121 gcacgagagc ctgatcctgc tgagcgaaaa cctgcgtaaa gatgtggagg ctgtgaccgg
     3181 cagcccggct agccagacca gcatttgcat cggtattctg ctgcgcagcg ccgagctggc
     3241 gctgctgctg cacccagtgg accaggcgaa caccctgaag agcccggtga gcgaaagcgt
     3301 gagcccagtg gtgccagatt acctgccgac cgagaacggt gacttcctga gcagcaagcg
     3361 taaacagatt agccgcgata tcaaccgtat tcgcagcgtg accgtgaacc acatgagcga
     3421 taaccgtagc atgagcgtgg acctgagcca catcccgctg aaggacccgc tgctgttcaa
     3481 aagcgccagc gacaccaacc tgcagaaagg catcagcttt atggattatc tgagcgacaa
     3541 gcacctgggc aaaattagcg aggacgaaag cagcggtctg gtgtacaaga gcggcagcgg
     3601 cgagatcggt agcgaaacca gcgacaagaa agatagcttc tataccgata gcagcagcat
     3661 cctgaactac cgcgaggaca gcaacattct gagctttgac agcgatggca accagaacat
     3721 tctgagcagc accctgacca gcaagggtaa cgaaaccatc gaaagcattt tcaaggctga
     3781 ggatctgctg ccggaagctg ccagcctgag cgaaaacctg gacatcagca aagaggaaac
     3841 cccgccggtg cgtaccctga agagccagag cagcctgagc ggcaagccga aagagcgttg
     3901 cccaccaaac ctggctccac tgtgcgtgag ctataagaac atgaaacgta gcagcagcca
     3961 gatgagcctg gataccatca gcctggacag catgattctg gaggaacagc tgctggaaag
     4021 cgacggcagc gatagccaca tgttcctgga gaagggtaac aagaaaaaca gcaccaccaa
     4081 ctatcgtggt accgctgaaa gcgtgaacgc tggtgctaac ctgcagaact acggtgaaac
     4141 cagcccggac gccattagca ccaacagcga gggcgcgcag gaaaaccacg acgatctgat
     4201 gagcgtggtg gtgtttaaga tcaccggcgt gaacggcgag atcgatattc gtggcgagga
     4261 caccgaaatc tgcctgcagg tgaatcaggt gaccccggat cagctgggca acattagcct
     4321 gcgtcactac ctgtgcaacc gtccagtggg tagcgaccag aaggccgtga tccacagcaa
     4381 aagcagcccg gagattagcc tgcgtttcga aagcggtcca ggtgctgtga tccacagcct
     4441 gctggccgag aaaaacggtt ttctgcagtg ccacattgag aactttagca ccgaattcct
     4501 gaccagcagc ctgatgaaca tccagcactt cctggaggac gaaaccgtgg cgaccgtgat
     4561 gccgatgaag atccaggtga gcaacaccaa gattaacctg aaagacgata gcccgcgtag
     4621 cagcaccgtg agcctggaac cagctccagt gaccgtgcac attgatcacc tggtggtgga
     4681 gcgtagcgac gatggcagct ttcacatccg cgacagccac atgctgaaca ccggtaacga
     4741 tctgaaggag aacgtgaaaa gcgacagcgt gctgctgacc agcggcaaat acgatctgaa
     4801 gaaacagcgt agcgtgaccc aggctaccca gaccagcccg ggtgtgccgt ggccaagcca
     4861 gagcgccaac ttcccggaat tcagctttga cttcacccgc gagcagctga tggaggaaaa
     4921 cgagagcctg aagcaggaac tggctaaggc caaaatggcg ctggctgagg cacacctgga
     4981 aaaagacgct ctgctgcacc acattaagaa gatgaccgtt gagggtggat ctggaggtcg
     5041 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcatggtga gcaagggcga
     5101 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     5161 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     5221 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     5281 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     5341 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     5401 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     5461 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     5521 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     5581 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     5641 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     5701 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     5761 ctccaccggc ggcatggacg agctgtacaa gtaagcggcc gcgactctag atcataatca
     5821 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     5881 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     5941 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6001 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     6061 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6121 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6181 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6241 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6301 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6361 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6421 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6481 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6541 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6601 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     6661 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6721 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6781 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6841 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6901 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6961 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     7021 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     7081 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     7141 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     7201 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     7261 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     7321 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     7381 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     7441 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     7501 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     7561 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     7621 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     7681 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7741 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7801 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     7861 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     7921 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     7981 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     8041 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     8101 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     8161 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     8221 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     8281 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8341 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8401 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8461 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8521 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8581 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8641 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8701 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8761 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8821 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8881 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8941 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9001 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     9061 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     9121 ccatgcat
//
