LOCUS       sequence-371648-        6628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mammalian expression plasmid encoding miRFP-tagged human OCT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  AUTHORS   Chiu DC, Baskin JM
  TITLE     Organelle-Selective Membrane Labeling through Phospholipase 
            D-Mediated Transphosphatidylation.
  JOURNAL   JACS Au. 2022 Nov 28;2(12):2703-2713. doi: 10.1021/jacsau.2c00419. 
            eCollection 2022 Dec 26.
  PUBMED    36590261
REFERENCE   2  (bases 1 to 6628)
  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"; doi: 
            "10.1021/jacsau.2c00419"; journalName: "JACS Au"; date: "2022-11-28-
            28"; volume: "2"; issue: "12"; pages: "2703-2713"
COMMENT     Sequence Label: hOCT1-miRFP
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        410..613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2386..2395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2392..3339
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    3461..3582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3552..3571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3589..4044)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3726..3747)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3938..3957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4071..4175
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4177..4534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4177..4368
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4200..4220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4385..4520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4569..5363
                     /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             4569..5363
                     /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(4623..4642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5233..5252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5551..5570)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5595..5642
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5971..6559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6460..6479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
       61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      601 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      661 gaggccacca tgcccaccgt ggacgatatc ctggagcaag tgggcgagag cggctggttc
      721 cagaagcagg cctttctgat cctgtgcctg ctgtccgccg ccttcgcccc tatctgcgtg
      781 ggcatcgtgt tcctgggctt taccccagac caccactgcc agagcccagg agtggcagag
      841 ctgtctcaga gatgtggctg gagcccagcc gaggagctga actacacagt gccaggactg
      901 ggacctgcag gagaggcctt tctgggccag tgccggagat atgaggtgga ctggaaccag
      961 tctgccctga gctgcgtgga tcctctggcc tccctggcca ccaataggtc tcacctgcca
     1021 ctgggaccat gccaggacgg atgggtgtac gataccccag gcagctccat cgtgacagag
     1081 ttcaacctgg tgtgcgccga ctcttggaag ctggatctgt ttcagagctg tctgaatgcc
     1141 ggcttcctgt ttggcagcct gggcgtgggc tatttcgccg atagatttgg caggaagctg
     1201 tgcctgctgg gaaccgtgct ggtgaacgcc gtgtccggcg tgctgatggc cttcagcccc
     1261 aattacatgt ccatgctgct gtttcgcctg ctgcagggcc tggtgtctaa gggcaattgg
     1321 atggccggct ataccctgat cacagagttc gtgggatccg gatctaggag gacagtggcc
     1381 atcatgtacc agatggcctt taccgtggga ctggtggccc tgacaggact ggcatatgca
     1441 ctgccacact ggaggtggct gcagctggcc gtgtccctgc ccaccttcct gtttctgctg
     1501 tactattggt gcgtgcctga gagcccaaga tggctgctgt cccagaagag gaacacagag
     1561 gccatcaaga tcatggacca catcgcccag aagaatggca agctgccccc tgccgacctg
     1621 aagatgctga gcctggagga ggatgtgacc gagaagctga gcccctcctt cgccgacctg
     1681 tttcggaccc ctcggctgag aaagaggaca ttcatcctga tgtacctgtg gtttaccgat
     1741 tccgtgctgt atcagggcct gatcctgcac atgggcgcca catccggcaa cctgtacctg
     1801 gacttcctgt attctgccct ggtggagatc ccaggcgcct ttatcgccct gatcacaatc
     1861 gatcgcgtgg gccggatcta ccccatggca atgtctaacc tgctggcagg agcagcatgt
     1921 ctggtcatga tcttcatcag ccccgacctg cactggctga atatcatcat catgtgcgtg
     1981 ggccgcatgg gcatcaccat cgccatccag atgatctgtc tggtgaacgc cgagctgtat
     2041 cctacatttg tgcggaatct gggcgtgatg gtgtgctcta gcctgtgcga tatcggcggc
     2101 atcatcaccc ctttcatcgt gtttcgcctg agggaagtgt ggcaggccct gccactgatc
     2161 ctgttcgccg tgctgggact gctggcagca ggagtgaccc tgctgctgcc tgagacaaag
     2221 ggcgtggccc tgccagagac catgaaggat gccgagaacc tgggcagaaa ggccaagccc
     2281 aaggagaata caatctacct gaaggtgcag acctccgagc cttctggcac aggaggctcc
     2341 ggaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtagca
     2401 ggtcatgcct ctggcagccc cgcattcggg accgcctctc attcgaattg cgaacatgaa
     2461 gagatccacc tcgccggctc gatccagccg catggcgcgc ttctggtcgt cagcgaacat
     2521 gatcatcgcg tcatccaggc cagcgccaac gccgcggaat ttctgaatct cggaagcgta
     2581 ctcggcgttc cgctcgccga gatcgacggc gatctgttga tcaagatcct gccgcatctc
     2641 gatcccaccg ccgaaggcat gccggtcgcg gtgcgctgcc ggatcggcaa tccctctacg
     2701 gagtactgcg gtctgatgca tcggcctccg gaaggcgggc tgatcatcga actcgaacgt
     2761 gccggcccgt cgatcgatct gtcaggcacg ctggcgccgg cgctggagcg gatccgcacg
     2821 gcgggttcac tgcgcgcgct gtgcgatgac accgtgctgc tgtttcagca gtgcaccggc
     2881 tacgaccggg tgatggtgta tcgtttcgat gagcaaggcc acggcctggt attctccgag
     2941 tgccatgtgc ctgggctcga atcctatttc ggcaaccgct atccgtcgtc gactgtcccg
     3001 cagatggcgc ggcagctgta cgtgcggcag cgcgtccgcg tgctggtcga cgtcacctat
     3061 cagccggtgc cgctggagcc gcggctgtcg ccgctgaccg ggcgcgatct cgacatgtcg
     3121 ggctgcttcc tgcgctcgat gtcgccgtgc catctgcagt tcctgaagga catgggcgtg
     3181 cgcgccaccc tggcggtgtc gctggtggtc ggcggcaagc tgtggggcct ggttgtctgt
     3241 caccattatc tgccgcgctt catccgtttc gagctgcggg cgatctgcaa acggctcgcc
     3301 gaaaggatcg cgacgcggat caccgcgctt gagagctaag cggccgcgac tctagatcat
     3361 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3421 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3601 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3661 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3721 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3781 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3841 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3901 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3961 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4021 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4201 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4261 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4381 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4441 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4501 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4561 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4621 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4681 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4741 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4801 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4861 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4921 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4981 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5041 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5101 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5161 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5221 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5281 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5341 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5401 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5521 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5581 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5641 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5701 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5761 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6601 ctggcctttt gctcacatgt tctttcct
//
