LOCUS       sequence-434118-        7520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpresses ORP6 with a neonGreen fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  AUTHORS   Doyle CP, Rectenwald A, Timple L, Hammond GRV
  TITLE     Orthogonal Targeting of SAC1 to Mitochondria Implicates ORP2 as a 
            Major Player in PM PI4P Turnover.
  JOURNAL   Contact (Thousand Oaks). 2024 Feb 7;7:25152564241229272. doi: 
            10.1177/25152564241229272. eCollection 2024 Jan-Dec.
  PUBMED    38327560
REFERENCE   2  (bases 1 to 7520)
  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: "Contact 
            (Thousand Oaks)."; date: "2024-02-7"; volume: "7:25152564241229272. 
            doi"; pages: " 10.1177/25152564241229272. eCollection 2024 Jan-Dec"
COMMENT     Sequence Label: NeonGreen-C1-ORP6
FEATURES             Location/Qualifiers
     source          1..7520
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4308..4429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4345..4364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4399..4418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4436..4891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4573..4594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4785..4804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5024..5381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5024..5215
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5047..5067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5232..5367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5416..6210
                     /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             5416..6210
                     /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(5470..5489)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6080..6099
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6398..6417)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6442..6489
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6818..7406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7307..7326
                     /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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
      841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ataccaagtc
      901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
      961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1321 tccggactca gatctatgag ttcagatgag aagggcattt cccctgctca taaaacatcc
     1381 actccaaccc atagaagtgc ctcctcttca acatcctccc aaagggacag taggcagagt
     1441 attcacatac tggagaggac tgcttcctct agcaccgagc cctctgtaag tcggcaattg
     1501 ctagaaccgg agccagtccc cctctccaag gaagctgaca gctgggaaat tatagaaggg
     1561 ctgaaaatag gccaaaccaa tgtccagaaa ccagacaaac atgagggctt tatgctgaag
     1621 aaaagaaaat ggcctttaaa aggctggcac aagcgttttt ttgtcctgga taatggaatg
     1681 ttaaagtatt caaaggcacc actcgatatt cagaaaggaa aggtccatgg gagcatagat
     1741 gtgggactct cagtcatgtc aattaaaaag aaagctcgaa gaatagacct tgacaccgaa
     1801 gagcacatct atcatttgaa ggtgaaatcc caggactggt ttgatgcatg ggtctccaaa
     1861 ctgcgacatc atcggttgta tcgtcagaat gaaattgtga gatcaccaag agatgctagt
     1921 tttcacatat ttccttcaac gtccacagct gaatcctcac cagctgctaa tgtttctgta
     1981 atggatggaa agatgcaacc aaacagcttt ccgtggcagt cccctttacc atgcagcaat
     2041 agcctccctg caacgtgcac aactggccag agtaaagtgg cagcctggtt acaggactcg
     2101 gaagagatgg acaggtgtgc agaagatctt gcacattgcc agtcaaacct tgtggaactt
     2161 agcaaactcc tgcaaaattt ggaaatactt cagagaactc agtcggcacc taactttact
     2221 gacatgcagg ctaactgtgt agatatttca aagaaagaca agcgggtcac aagacgatgg
     2281 agaacaaaaa gtgtcagcaa agatacaaaa atacaactgc aggttccttt cagtgctacc
     2341 atgtcaccag ttcgcttgca ttcctccaac cccaaccttt gtgcagatat tgaatttcag
     2401 actcccccta gccacctcac tgaccctctg gaaagttcaa cagattatac aaagctgcaa
     2461 gaagaatttt gtctaatcgc acagaaagtg cattctcttt tgaagtctgc atttaatagc
     2521 atagctatag agaaggagaa gctgaagcag atggtttccg agcaggatca cagtaaaggc
     2581 cacagcacgc agatggcacg gctccgacag tcactgtctc aggcactcaa ccagaatgct
     2641 gaactaagga gtcggttgaa cagaatacat tcagagtcta ttatttgtga tcaggttgtc
     2701 agtgtaaata ttattcctag ccctgatgag gctggtgagc aaatccatgt cagtctcccc
     2761 ttatcacagc aagtagccaa tgagagccgc ctctccatgt cagagtctgt ttctgagttc
     2821 tttgatgccc aagaggtgct cctctctgca agttcgtcag agaatgaggc ttcagatgat
     2881 gagtcttaca tcagtgatgt gagtgataat atatctgaag acaacaccag tgttgcagac
     2941 aatatttctc ggcaaatcct gaatggggag cttacaggag gggccttccg aaatgggcgt
     3001 cgagcatgcc tgccagctcc ttgtcctgac accagtaaca ttaacctgtg gaatatcttg
     3061 aggaacaaca ttggtaaaga cctgtctaaa gtctctatgc ctgtggagct aaacgagccg
     3121 ctcaacaccc tgcagcacct ctgtgaggaa atggaataca gcgagctcct ggacaaggct
     3181 tcggaaactg atgatccata tgagcgcatg gttctcgttg ccgcatttgc agtttcagga
     3241 tactgctcca cctatttcag agcaggaagt aagccattca acccagtcct tggggagact
     3301 tatgaatgca ttagagaaga caagggattc cgctttttct cagaacaggt tagccatcat
     3361 ccacccattt ctgcctgtca ctgtgaatca aagaattttg tgttttggca agatatcaga
     3421 tggaaaaaca agttctgggg gaagtcgatg gaaatcctgc ctgttggaac actgaatgtc
     3481 atgcttccaa agtatggaga ttactatgtg tggaataaag tcaccacttg catacacaac
     3541 atcctcagtg ggagaagatg gatagaacat tatggagaag taaccatcag aaataccaaa
     3601 agcagtgttt gcatttgcaa actcacattt gtcaaggtga attattggaa ttctaacatg
     3661 aatgaagtcc agggggtggt gatagatcag gaggggaagg cggtgtaccg gctgtttgga
     3721 aagtggcatg aaggactcta ctgtggtgtg gccccctctg caaagtgcat ttggagacca
     3781 ggttccatgc caacaaacta tgagctgtac tatggcttca caaggtttgc tattgagctc
     3841 aatgagttag atccagtact aaaagatctc cttccaccaa cagacgcccg gttccggcca
     3901 gatcaaagat ttttggaaga aggaaattta gaagctgcag catcagagaa gcaaagagta
     3961 gaggaactcc agagatctcg gagacgatat atggaagaaa acaatcttga acatatacca
     4021 aaatttttta aaaaagttat tgatgccaat caaagagaag cctgggtttc taacgacacc
     4081 tactgggagc ttcgaaagga ccctgggttt agcaaagtag acagccctgt tctttggtag
     4141 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
     4201 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4261 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     4441 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4501 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4561 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4621 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4681 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4741 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4801 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4861 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     5041 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5101 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5221 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5281 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5341 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     5401 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5461 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5521 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5581 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5641 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5701 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5761 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5821 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5881 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5941 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6001 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6061 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6121 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6181 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6241 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6301 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6361 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     6421 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     6481 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6541 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6601 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6661 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6961 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7441 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7501 accgtattac cgccatgcat
//
