LOCUS       sequence-489488-        9140 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  For exogenous expression of codon optimized BLTP3A (UHRF1BP1) 
            chimera-2 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 9140)
  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 9140)
  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-2
FEATURES             Location/Qualifiers
     source          1..9140
                     /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             5096..5806
                     /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(5241..5259)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5509..5528
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5928..6049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5965..5984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6019..6038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6056..6511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6193..6214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6405..6424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6538..6642
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6644..7001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6644..6835
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6667..6687)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6852..6987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7036..7830
                     /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             7036..7830
                     /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(7090..7109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7700..7719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8018..8037)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8062..8109
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8438..9026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8927..8946
                     /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 catggccggc atcatcaaaa aacaaatcct
      661 gaaacacctg tcccgcttta ccaagaacct gtccccagat aagatcaacc tgtccaccct
      721 gaagggcgag ggtgaactga aaaacctgga gctggatgag gaagtgctgc agaacatgct
      781 ggacctgccg acctggctgg ctatcaacaa ggtgttctgc aacaaagcca gcatccgtat
      841 tccgtggacc aagctgaaaa cccacccgat ctgcctgagc ctggacaaag tgattatgga
      901 aatgagcacc tgcgaggaac cacgcagccc gaacggtcca agcccaattg ctaccgctag
      961 cggtcagagc gagtatggtt ttgcggagaa agtggtggaa ggcatcttca 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 caccacctgg ttatcagcca
     1621 cctggatctg cacatctgcg acgatattca cgcgaaggag aaagaaagca accgtcgcat
     1681 caccggcggt gctatgcagc tgagcttcac ccagctgacc attgattact atccgtatca
     1741 caaagcgggc gacagctgca accactggat gtacttcagc gacgcgacca agaccaaaaa
     1801 cggttgggct aacgaactgc tgcacgagtt tgaatgcaac gtggagatgc tgaagcaggc
     1861 ggtgaaagat cacaacgtgg gcagcccgcc gaagagccca acccacgcta gcccacagca
     1921 cacccagacc gagaaggact atccgctgaa aggtacctgc cgtaccccga gcgtgctgag
     1981 ccagcagagc aaggccaaac tgatgagcag cagcgtggtg gtgcgtctgg cggacttcaa
     2041 catctaccag gtgagcaccg ctgaacagtg ccgcagcagc ccgaaaagca tgatctgctg
     2101 caacaagaaa agcctgtatc tgccgcagga gatgagcgcg gtgtacatcg agttcaccga
     2161 atactattac ccggacggca aggattttcc gattccgagc ccgaacctgt atagccagct
     2221 gaacgctctg cagttcaccg tggatgaacg tagcatcctg tggctgaacc agttcctgct
     2281 ggacctgaag cagagcctga accagtttat ggccgtgtac aagctgaacg ataacagcaa
     2341 aagcgacgag cacgtggatg tgcgtgtgga cggtctgatg ctgaaatttg tgattccgag
     2401 cgaggtgaag agcgaatgcc accaggacca gccacgtgcc attagcatcc agagcagcga
     2461 aatgatcgcg accaacaccc gtcactgccc aaactgccgt cacagcgatc tggaggcgct
     2521 gtttcaggac ttcaaggact gcgatttctt tagcaaaacc tataccagct ttccgaagag
     2581 ctgcgataac ttcaacctgc tgcacccgat ctttcagcgt cacgcccacg agcaggacac
     2641 caagatgcac gaaatctaca aaggcaacat taccccgcag ctgaacaaga acaccctgaa
     2701 aaccagcgcc gctaccgacg tgtgggccgt gtacttcagc cagttctgga tcgactacga
     2761 gggtatgaag agcggcaaag gtcgtccgat tagctttgtg gatagcttcc cgctgagcat
     2821 ttggatctgc cagccgaccc gctatgccga gagccagaag gaaccgcaga cctgcaacca
     2881 ggtgagcctg aacaccagcc agagcgaaag cagcgacctg gctggccgtc tgaagcgcaa
     2941 gaaactgctg aaagagtatt acagcaccga gagcgaaccg ctgaccaacg gcggtcagaa
     3001 accgagcagc agcgatacct tctttcgttt cagcccgagc agcagcgaag ccgacatcca
     3061 cctgctggtg cacgtgcaca agcacgtgag catgcagatt aaccactatc agtacctgct
     3121 gctgctgttt ctgcacgaga gcctgatcct gctgagcgaa aacctgcgta aagatgtgga
     3181 ggctgtgacc ggcagcccgg ctagccagac cagcatttgc atcggtattc tgctgcgcag
     3241 cgccgagctg gcgctgctgc tgcacccagt ggaccaggcg aacaccctga agagcccggt
     3301 gagcgaaagc gtgagcccag tggtgccaga ttacctgccg accgagaacg gtgacttcct
     3361 gagcagcaag cgtaaacaga ttagccgcga tatcaaccgt attcgcagcg tgaccgtgaa
     3421 ccacatgagc gataaccgta gcatgagcgt ggacctgagc cacatcccgc tgaaggaccc
     3481 gctgctgttc aaaagcgcca gcgacaccaa cctgcagaaa ggcatcagct ttatggatta
     3541 tctgagcgac aagcacctgg gcaaaattag cgaggacgaa agcagcggtc tggtgtacaa
     3601 gagcggcagc ggcgagatcg gtagcgaaac cagcgacaag aaagatagct tctataccga
     3661 tagcagcagc atcctgaact accgcgagga cagcaacatt ctgagctttg acagcgatgg
     3721 caaccagaac attctgagca gcaccctgac cagcaagggt aacgaaacca tcgaaagcat
     3781 tttcaaggct gaggatctgc tgccggaagc tgccagcctg agcgaaaacc tggacatcag
     3841 caaagaggaa accccgccgg tgcgtaccct gaagagccag agcagcctga gcggcaagcc
     3901 gaaagagcgt tgcccaccaa acctggctcc actgtgcgtg agctataaga acatgaaacg
     3961 tagcagcagc cagatgagcc tggataccat cagcctggac agcatgattc tggaggaaca
     4021 gctgctggaa agcgacggca gcgatagcca catgttcctg gagaagggta acaagaaaaa
     4081 cagcaccacc aactatcgtg gtaccgctga aagcgtgaac gctggtgcta acctgcagaa
     4141 ctacggtgaa accagcccgg acgccattag caccaacagc gagggcgcgc aggaaaacca
     4201 cgacgatctg atgagcgtgg tggtgtttaa gatcaccggc gtgaacggcg agatcgatat
     4261 tcgtggcgag gacaccgaaa tctgcctgca ggtgaatcag gtgaccccgg atcagctggg
     4321 caacattagc ctgcgtcact acctgtgcaa ccgtccagtg ggtagcgacc agaaggccgt
     4381 gatccacagc aaaagcagcc cggagattag cctgcgtttc gaaagcggtc caggtgctgt
     4441 gatccacagc ctgctggccg agaaaaacgg ttttctgcag tgccacattg agaactttag
     4501 caccgaattc ctgaccagca gcctgatgaa catccagcac ttcctggagg acgaaaccgt
     4561 ggcgaccgtg atgccgatga agatccaggt gagcaacacc aagattaacc tgaaagacga
     4621 tagcccgcgt agcagcaccg tgagcctgga accagctcca gtgaccgtgc acattgatca
     4681 cctggtggtg gagcgtagcg acgatggcag ctttcacatc cgcgacagcc acatgctgaa
     4741 caccggtaac gatctgaagg agaacgtgaa aagcgacagc gtgctgctga ccagcggcaa
     4801 atacgatctg aagaaacagc gtagcgtgac ccaggctacc cagaccagcc cgggtgtgcc
     4861 gtggccaagc cagagcgcca acttcccgga attcagcttt gacttcaccc gcgagcagct
     4921 gatggaggaa aacgagagcc tgaagcagga actggctaag gccaaaatgg cgctggctga
     4981 ggcacacctg gaaaaagacg ctctgctgca ccacattaag aagatgaccg ttgagggtgg
     5041 atctggaggt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcatggt
     5101 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     5161 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     5221 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     5281 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     5341 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     5401 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     5461 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     5521 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     5581 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     5641 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     5701 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     5761 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagcgg ccgcgactct
     5821 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5881 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     6061 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6241 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6481 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     6541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     6661 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6721 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6841 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6901 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6961 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     7021 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     7081 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     7141 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     7201 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     7261 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     7321 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     7381 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     7441 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     7501 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     7561 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     7621 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     7681 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     7741 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7801 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     7861 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     7921 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     7981 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     8041 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     8101 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     8161 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     8221 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     8281 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     8341 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8401 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8581 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8821 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     9001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     9061 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     9121 accgtattac cgccatgcat
//
