LOCUS       sequence-219937-        7918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of OCTN2 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7918)
  AUTHORS   Jurkiewicz D, Michalec K, Skowronek K, Nalecz KA
  TITLE     Tight junction protein ZO-1 controls organic cation/carnitine 
            transporter OCTN2 (SLC22A5) in a protein kinase C-dependent way.
  JOURNAL   Biochim Biophys Acta. 2017 May;1864(5):797-805. doi: 
            10.1016/j.bbamcr.2017.02.014. Epub 2017 Feb 28.
  PUBMED    28257821
REFERENCE   2  (bases 1 to 7918)
  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: 
            "10.1016/j.bbamcr.2017.02.014"; journalName: "Biochim Biophys Acta";
            date: "2017-05"; volume: "1864"; issue: "5"; pages: "797-805"
COMMENT     Sequence Label: p3 FLAG_CMV14/OCTN2 (delta554_557/PDZ)
FEATURES             Location/Qualifiers
     source          1..7918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2284..2349
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2360..2982
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2559..2578)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3006..3026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3024..3321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3375..4169
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3429..3448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4039..4058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4821..4955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4858..4877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4912..4931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4960..4979)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4961..4979)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4993..5009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4993..5009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5006..5028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5017..5033
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5041..5047,5048..5065,5066..5071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5086..5107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5224..5241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5395..5983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5475..5494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6154..6945,6946..7014))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6777..6796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7015..7119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7146..7601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7233..7252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7443..7464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7727..7749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7741..7758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7742..7758
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaagc ttatgcggga ctacgacgag gtgaccgcct tcctgggcga gtgggggccc
      661 ttccagcgcc tcattttctt cctgctcagc gccagcatca tccccaatgg cttcaatggt
      721 atgtctatcg tgttcctggc ggggaccccg gagcaccgtt gccttgtgcc tcacaccgtg
      781 aacttgagca gcgcgtggcg caaccacagt atcccgttgg agacgaagga cggacgacag
      841 gtgcctcaga gctgccgccg ctaccgactg gccaccatcg ccaacttctc tgcgctgggg
      901 ctggagccgg gacgggacgt ggacctggag cagctggagc aagagaactg cctggatggc
      961 tgggagtaca acaaggacgt cttcctgtcc accatcgtga cagagtggga cctggtgtgt
     1021 aaggatgact ggaaagcccc actcaccacc tccttgtttt tcgtgggtgt gctgatgggc
     1081 tccttcattt cgggacagct ctcagacagg tttggtcgca agaatgtgct gtttttgacc
     1141 atgggcatgc agactggctt cagcttcctg cagctcttct ctgtgaactt cgagatgttt
     1201 acagtgcttt ttgtccttgt tggcatgggt cagatctcca actacgtggc agcatttgtc
     1261 ctgggaacag aaattctttc caagtcaatt cgaattatat tcgccacctt aggagtttgc
     1321 atattttatg catttggctt catggtgctg cctctgtttg catacttcat cagagactgg
     1381 aggatgctgc tgcttgcgct cactgtacca ggggtgctgt gtggggctct ctggtggttc
     1441 atccctgagt ccccccgatg gctcatctct caaggccgag ttaaagaggc agaggtgatc
     1501 atccgcaaag ctgccaaatt caatgggata gttgcgcctt ccactatctt cgatcccagt
     1561 gagttacaag acttaaattc caagaagcct cagtcgcacc acatttatga tctggtccga
     1621 acacgaaata tcaggatcat caccatcatg tccataatcc tgtggctgac catatcagtg
     1681 ggctacttcg gactttctct tgacactcct aacttgcatg gggacatcta tgtgaactgc
     1741 ttcctactgg cagctgttga agtccccgcc tatgtgttgg cctggctgtt gctacagcac
     1801 ctgccccggc ggtattctat ctctgctgcc ctcttcctgg gtggcagtgt ccttctcttc
     1861 atacagctgg tgccttcaga attgttttac ttgtccactg ccctggtgat ggtgggcaag
     1921 tttggaatca cctctgccta ttccatggtc tatgtgtaca cggccgagct gtaccccact
     1981 gtggtcagaa acatgggtgt gggggtcagc tccacagcat cccgccttgg cagcatcctg
     2041 tctccctact ttgtctacct cggtgcctat gatcgcttcc tgccttatat cctcatggga
     2101 agtctgacca tcctgacagc tatcctcacc ttgttcttcc cagagagctt tggtgcccct
     2161 ctccccgaca ccattgacca gatgctcagg gtcaaaggaa taaaacaatg gcaaatccaa
     2221 agccaaacaa ggacgcaaaa agatggtgga gaaagcccaa cggtcctaaa gggatcccgg
     2281 gctgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     2341 gatgacaagt agtgatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2401 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2461 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2521 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2581 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2641 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2701 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2761 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2821 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     2881 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     2941 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     3001 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3061 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3181 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3241 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3301 gtagtgagga ggcttttttg gaggaattga tcagcttggg atctgatcaa gagacaggat
     3361 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3421 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3481 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3541 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3601 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3661 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3721 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3781 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3841 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3901 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3961 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4021 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4081 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4141 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4201 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4261 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4321 gctggagttc ttcgcccacc ccgggctcga tcccctcgcg agttggttca gctgctgcct
     4381 gaggctggac gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca tccaggaaac
     4441 cagcagcggc tatccgcgca tccatgcccc cgaactgcag gagtggggag gcacgatggc
     4501 cgctttggtc gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg
     4561 catctcatga gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg
     4621 cgcctgctgc gcgacggcga gctgctcacc acccactcgc caagctggaa ccgtaaaaag
     4681 gccgcgttgc tggcgttttt ccataggctc cgccgatcat aatcagccat accacatttg
     4741 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4801 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4861 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4921 ccaaactcat caatgtatct tatcatgtct ggatcaattc cctatagtga gtcgtattaa
     4981 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5101 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7141 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     7321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     7381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     7441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     7501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     7561 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     7621 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7681 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7741 tgtaaaacga cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc
     7801 atattagtca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     7861 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
