LOCUS       sequence-207196-        7559 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for codon optimized mutant hSERT with 
            internal MYC tag inserted in ECL2, mutation F465L conferring gain of
            function phenotype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   Young HJ, Chan M, Selvam B, Szymanski SK, Shukla D, Procko E
  TITLE     Deep Mutagenesis of a Transporter for Uptake of a Non-Native 
            Substrate Identifies Conformationally Dynamic Regions
  JOURNAL   bioRxiv 2021.04.19.440442
REFERENCE   2  (bases 1 to 7559)
  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: "bioRxiv 
            2021.04.19.440442"
COMMENT     Sequence Label: pCEP4-SERTF465L-myc
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     342..361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        872..1017
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             1060..2097
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(2095..2114)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2139..2187
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        2841..3220
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3221..3424
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3375..3395
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3421..3445
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3473..3482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4148..4177
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    5466..5600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5490..5509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5544..5563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5743..5765)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
       61 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      481 tgctggcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
      541 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
      601 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccccgag atgcgccgcg
      661 tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat
      721 tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga
      781 ggtgccgccc tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga
      841 agaaatatat ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc
      901 attggcgaat tcgaacacgc agatgcagtc ggggcggcgc ggtccgaggt ccacttcgca
      961 tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaacag
     1021 cgtcaacagc gtgccgcaga tcccgggggg caatgagata tgaaaaagcc tgaactcacc
     1081 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     1141 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     1201 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     1261 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     1321 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     1381 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     1441 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     1501 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     1561 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     1621 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     1681 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     1741 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     1801 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     1861 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     1921 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     1981 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     2041 agtggaaacc gacgccccag cactcgtccg gatcgggaga tgggggaggc taactgaaac
     2101 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     2161 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     2221 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     2281 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     2341 gccctgccat agccactggc cccgtgggtt agggacgggg tcccccatgg ggaatggttt
     2401 atggttcgtg ggggttatta ttttgggcgt tgcgtggggt caggtccacg actggactga
     2461 gcagacagac ccatggtttt tggatggcct gggcatggac cgcatgtact ggcgcgacac
     2521 gaacaccggg cgtctgtggc tgccaaacac ccccgacccc caaaaaccac cgcgcggatt
     2581 tctggcgtgc caagctagtc gaccaattct catgtttgac agcttatcat cgcagatccg
     2641 ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga tctatacatt
     2701 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     2761 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     2821 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     2881 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2941 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3001 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3061 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     3121 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     3181 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3241 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3301 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     3361 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     3421 agctcgttta gtgaaccgtc agatctctag aagctgggta ccagctgcta gcgccaccat
     3481 ggagaccacc cccctgaaca gtcaaaagca actctctgcc tgtgaggatg gagaggactg
     3541 tcaggagaat ggcgttcttc aaaaggttgt acccacccct ggagataagg tggaaagcgg
     3601 ccagatttcc aacggctaca gcgccgtgcc atcacctgga gccggggatg atacacggca
     3661 cagcattccc gcaaccacca cgacactggt tgcagagctt caccagggcg agagggagac
     3721 gtggggcaaa aaggtagatt ttctgctttc cgtgatcggt tacgcggtcg atctggggaa
     3781 cgtgtggagg ttcccttaca tttgttacca aaacggaggg ggtgccttct tgctgcccta
     3841 cacaattatg gcgatttttg ggggtatccc gctcttctac atggaactgg ccttgggtca
     3901 gtaccaccgg aacggctgca ttagcatttg gcgcaagata tgccccattt tcaagggaat
     3961 cggatacgca atctgtataa tcgcgttcta tattgcatca tattacaaca ccatcatggc
     4021 ctgggccctt tattacttga tcagcagttt caccgaccaa ttgccgtgga cgagctgtaa
     4081 gaactcatgg aataccggga actgtaccaa ctacttctct gaggacagct caggaggtag
     4141 cgggggtgaa cagaagctga ttagcgagga ggacttgggc agtccaggcg gaagcggcaa
     4201 catcacttgg acactccata gtacgagccc cgcagaggag ttttacacca ggcacgtact
     4261 gcagattcat aggagcaagg gtctgcaaga cctgggcggt attagctggc agctcgcatt
     4321 gtgcataatg ttgatcttta cggtgatcta tttctctatc tggaagggag taaagacatc
     4381 cggcaaggtg gtgtgggtga cggcgacctt cccctacatt atactcagcg tgttgttggt
     4441 gaggggggca accctgcctg gggcctggcg aggcgtgctg ttctatctca agccaaactg
     4501 gcagaagctt cttgaaaccg gagtgtggat cgacgcggcc gcgcaaatct tcttttccct
     4561 cggtcccggc ttcggggtcc ttctggcatt cgcaagctac aacaagttca acaataactg
     4621 ctaccaggac gccctcgtga caagtgtagt gaactgcatg acctctttcg tgagcgggtt
     4681 tgtgatcttc acagtactcg gatatatggc tgagatgagg aacgaagatg ttagtgaggt
     4741 tgccaaagac gctggcccga gcctgttgtt catcacatac gccgaggcta tcgcgaacat
     4801 gcccgcatct actttcttcg ccatcatctt tttcctgatg ctgataaccc ttgggctgga
     4861 tagcaccttc gccggactgg agggtgtcat cactgccgtt ctcgacgagt tcccccatgt
     4921 gtgggccaag cgacgggaaa ggctcgtgct ggccgtcgta atcacatgct tcttcggctc
     4981 actggtgacc cttacctttg gcggtgcgta cgtggtcaag ctgctcgaag agtacgccac
     5041 tggaccggcg gttctgaccg tggcattgat cgaggccgtg gccgtaagct ggttttacgg
     5101 catcacgcag ttctgtaggg acgtgaaaga gatgctgggc ttttctcccg gttggttttg
     5161 gaggatttgt tgggtagcaa ttagccccct gtttctcctt ttcattattt gctcttttct
     5221 gatgtctcca ccgcagctcc gactgtttca gtacaactac ccctactggt caattatcct
     5281 gggctattgt atcggcacct caagtttcat ctgcatcccc acatacatcg catacagact
     5341 gataattacg cccggtactt ttaaggagcg aatcattaaa agcataaccc ccgaaacccc
     5401 caccgagatc ccttgtggcg atataagact caatgcggtc taatagctcg aggccggcaa
     5461 ggccggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5581 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     5641 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     5701 atgatccggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5761 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5821 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg aggtcgactc tagaggatcg
     5881 atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca
     5941 gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt
     6001 gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat
     6061 ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt
     6121 ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc
     6181 tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat
     6241 cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt
     6301 tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata
     6361 ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg
     6421 aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca
     6481 cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg
     6541 gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc
     6601 attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga
     6661 ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca
     6721 gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg caataaatac
     6781 tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca
     6841 agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc
     6901 tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga
     6961 accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg
     7021 actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc
     7081 cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca
     7141 gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga
     7201 ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc
     7261 cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct
     7321 ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata
     7381 ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca
     7441 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     7501 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaat
//
