LOCUS       sequence-434116-        7286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpresses ORP3 with a neonGreen Fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  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 7286)
  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-ORP3
FEATURES             Location/Qualifiers
     source          1..7286
                     /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    4074..4195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4111..4130)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4165..4184
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4202..4657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4339..4360)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4551..4570
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4684..4788
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4790..5147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4790..4981
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4813..4833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4998..5133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5182..5976
                     /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             5182..5976
                     /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(5236..5255)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5846..5865
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6164..6183)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6208..6255
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6584..7172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7073..7092
                     /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 gatctatgat gagtgatgag aagaaccttg gtgtgtccca aaaattggta
     1381 tcaccttcaa ggagcacaag tagctgctct tccaagcaag gaagtcgaca ggacagctgg
     1441 gaagtggtgg aaggactgag gggggagatg aattacaccc aggagccacc agttcagaaa
     1501 ggatttttgc tgaaaaagag gaagtggccc ttaaaaggct ggcataagag attcttctat
     1561 ctggacaaag gaatcttgaa atatgccaag agccaaaccg atatagagag agagaagctg
     1621 catggctgca ttgatgtcgg gctctcagtg atgtctgtaa agaagtcatc aaaatgcata
     1681 gaccttgaca ccgaggagca catctaccat ctgaaggtca agtcagaaga agtctttgat
     1741 gagtgggtat cgaaacttcg ccaccacaga atgtatcgtc agaatgaaat tgccatgttt
     1801 ccacatgaag ttaaccactt tttctcaggg tccaccatca cagactcttc atctggggtg
     1861 tttgactcca tttcaagtag gaagcgtagc agtatatcaa agcagaattt atttcaaact
     1921 ggaagcaatg tatcattttc ttgtggtggt gagacacgag ttccattatg gttacagtct
     1981 tcagaggaca tggaaaaatg ctccaaagac ctggcgcact gtcatgccta cctggtagaa
     2041 atgagccagc tcctgcaaag catggacgtc ctgcatcgga catactcggc accagctatc
     2101 aacgccatcc aggtcccgaa acctttttct ggcccagtaa gactacactc ctccaatcct
     2161 aatttgtcaa cactagattt tggagaagag aaaaattatt ctgatggctc tgaaacctca
     2221 tcagagtttt ctaaaatgca agaagatctg tgtcatattg cccataaagt ttacttcact
     2281 ttaaggtcag cttttaatat catgtcagcg gagagagaga aactgaagca gctgatggag
     2341 caggatgcct cctcctcccc gtctgctcag gtcattggtc tgaagaacgc cctgtcatcc
     2401 gccctagcac aaaacacaga tcttaaagaa cgcttacgca gaatccatgc cgagtctctg
     2461 ctcctcgact cccccgctgt cgccaagtcg ggtgacaatc tggcagagga aaactccaga
     2521 gatgaaaacc gagctctagt tcatcagctt tctaatgaaa gtagactctc catcactgac
     2581 tccctttctg agttttttga tgctcaggaa gttctgttat ctccaagctc ttcagaaaac
     2641 gagatttctg atgatgactc atatgtcagt gacataagtg ataatctttc cttagataat
     2701 ctcagtaatg atttagataa tgagagacag accttggggc ctgtccttga tagtggtcgg
     2761 gaagcgaagt cccggagaag aacgtgcctg ccggcgccct gcccgagcag cagtaacatc
     2821 agcctgtgga acatcctgag gaacaacatc gggaaggacc tgtccaaggt ggccatgccg
     2881 gtggagctga acgagcccct gaacacgctg cagaggctct gcgaggagct ggagtacagc
     2941 gagctcctgg acaaggccgc gcagattccc agccccctgg aaaggatggt atatgtggca
     3001 gcctttgcca tatcagcgta tgcatctagc tactaccgag ctggaagcaa gccatttaat
     3061 ccggttcttg gagaaacata tgaatgtatt cgggaggaca agggcttcca gtttttttca
     3121 gaacaggtca gccaccatcc gcctatctct gcgtgtcatg ctgagtctag aaattttgtt
     3181 ttctggcaag atgtgagatg gaaaaacaaa ttctggggca aatccatgga aattgttcca
     3241 attggcacaa cccatgtgac tctgccagtt tttggggatc attttgagtg gaacaaagtg
     3301 acctcttgca tccataacat cttaagcggg cagaggtgga ttgagcacta tggagagatt
     3361 gtcatcaaga acctgcatga tgattcctgc tactgcaaag tgaattttat aaaggcaaaa
     3421 tactggagca ctaatgccca tgagattgaa ggcacagtgt ttgacaggag tggaaaagcg
     3481 gttcatcggc tgtttgggaa atggcatgaa agcatctact gtggcggcgg ctcctcttct
     3541 gcctgtgtat ggagagcaaa tcctatgccg aaaggctacg agcaatacta tagcttcaca
     3601 cagtttgcgc tggaattaaa tgaaatggat ccatcatcaa agtctttatt gccacctact
     3661 gacactcgat ttaggccaga ccagaggttt ctagaggaag ggaacttaga agaagctgaa
     3721 atacaaaagc agaggattga acaactgcag agagaaaggc ggcgggtctt agaagaaaat
     3781 catgtggagc accagcctcg gtttttcagg aaatccgacg atgactcttg ggtgagcaac
     3841 ggcacctatt tggaacttag aaaagatctt ggtttttcca aactggacca tcctgtctta
     3901 tggtgaaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     3961 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4021 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     4201 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4261 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4321 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4381 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4441 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4501 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4561 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4621 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4681 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4741 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4801 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4861 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4921 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4981 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5041 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5101 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     5161 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5221 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5281 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5341 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5401 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5461 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5521 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5581 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5641 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5701 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5761 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5821 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5881 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5941 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6001 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6061 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6121 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     6181 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     6241 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     6301 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     6361 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     6421 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6481 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6541 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6601 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6661 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6721 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6781 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6841 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6901 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6961 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7021 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7081 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7141 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7201 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7261 tggataaccg tattaccgcc atgcat
//
