LOCUS       sequence-434115-        6175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpresses ORP2 with a neonGreen fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  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 6175)
  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-ORP2
FEATURES             Location/Qualifiers
     source          1..6175
                     /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    2963..3084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3000..3019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3054..3073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3091..3546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3228..3249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3440..3459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3573..3677
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3679..4036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3679..3870
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3702..3722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3887..4022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4071..4865
                     /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             4071..4865
                     /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(4125..4144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4735..4754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5053..5072)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5097..5144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5473..6061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5962..5981
                     /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 gatctcgagc tcaaggcgga agcggtggga tgaacggaga ggaagaattc
     1381 tttgatgccg tcacaggctt tgattctgat aactcttctg gggaattttc agaggcaaat
     1441 cagaaagtca cgggaatgat tgacttagac accagcaaaa ataataggat tgggaaaact
     1501 ggggagaggc cctctcaaga gaacggaatt cagaaacaca ggacatcgct gccggctccc
     1561 atgttcagca gaagcgactt cagcgtgtgg accatcctga agaagtgtgt tggcctggag
     1621 ctgtccaaga tcacgatgcc aatcgccttc aacgagcctc tgagcttctt gcagcggatc
     1681 acggagtaca tggagcacgt gtacctcatc cacagggcct cctgccagcc ccagcccctg
     1741 gagaggatgc agtctgtggc tgcttttgct gtttcggctg tggcttccca gtgggagagg
     1801 accggcaaac catttaatcc actcttggga gaaacgtatg aattaatcag ggaagattta
     1861 ggattcagat ttatatcgga acaggtcagt caccaccccc ccatcagtgc gttccactcg
     1921 gaaggtctca accatgactt cctgttccat ggctccatct accccaagct caagttctgg
     1981 ggcaaaagcg tggaggcgga gccccgaggc accatcaccc tggagctgct caaacataat
     2041 gaagcctaca cctggaccaa ccccacctgc tgcgtccaca acgtcatcat cgggaagctg
     2101 tggatagagc agtatgggac agtggagatt ttaaaccaca gaactggaca taagtgtgtg
     2161 cttcacttta aaccgtgtgg attatttgga aaagaacttc acaaggtgga aggacacatt
     2221 caagacaaaa acaaaaagaa gctctttatg atctatggca aatggacgga atgtttgtgg
     2281 ggcatagatc ctgtttcgta tgaatccttc aagaagcagg agaggagagg tgaccacctg
     2341 agaaaggcca agctggatga agactccggg aaggctgaca gcgacgtggc tgacgacgtg
     2401 cctgtggccc aggagaccgt gcaggtcatt cctggcagca agctgctctg gaggatcaac
     2461 acccggcccc ccaactctgc ccagatgtat aatttcacca gtttcactgt gagcctcaac
     2521 gagctggaga caggcatgga gaagaccctg ccacccacgg actgccgcct gcgccctgac
     2581 atccgcggca tggagaatgg caacatggat ctggccagcc aggagaagga gcggctggag
     2641 gagaagcaga gagaagcacg gagggagcgg gccaaggagg aggcagagtg gcagacgagg
     2701 tggttctacc caggcaataa cccctacact gggacccccg actggttgta tgcaggggat
     2761 tactttgagc ggaatttctc cgactgccca gatatctact gagaattctg cagtcgacgg
     2821 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     2881 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2941 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3061 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     3121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3541 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3601 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3661 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3961 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4021 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4081 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4141 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4201 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4261 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4321 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4381 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4441 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4501 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4561 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4621 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4681 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4741 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4801 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4861 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4921 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4981 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5041 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5101 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5161 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5221 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5281 caacgtcggg gcggtaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6061 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
