LOCUS       sequence-205689-        9588 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for codon optimized mutant hSERT with 
            internal MYC tag inserted in ECL2, mutation C473H conferring gain of
            function phenotype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9588)
  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 9588)
  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-SERTC473H-myc
FEATURES             Location/Qualifiers
     source          1..9588
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(628..646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        714..818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(819..887,888..1679)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1037..1056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2339..2358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2869..3014
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3057..4094
                     /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(4092..4111)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4136..4184
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4838..5217
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5218..5421
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5372..5392
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5418..5442
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5470..5479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6145..6174
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    7463..7597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7487..7506)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7541..7560
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7740..7762)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ttggccctga tcctgagccg cccggggctc ctggtcttcc gcctcctcgt cctcgtcctc
       61 ttccccgtcc tcgtccatgg ttatcacccc ctcttctttg aggtccactg ccgccggagc
      121 cttctggtcc agatgtgtct cccttctctc ctaggccatt tccaggtcct gtacctggcc
      181 cctcgtcaga catgattcac actaaaagag atcaatagac atctttatta gacgacgctc
      241 agtgaataca gggagtgcag actcctgccc cctccaacag cccccccacc ctcatcccct
      301 tcatggtcgc tgtcagacag atccaggtct gaaaattccc catcctccga accatcctcg
      361 tcctcatcac caattactcg cagcccggaa aactcccgct gaacatcctc aagatttgcg
      421 tcctgagcct caagccaggc ctcaaattcc tcgtccccct ttttgctgga cggtagggat
      481 ggggattctc gggacccctc ctcttcctct tcaaggtcac cagacagaga tgctactggg
      541 gcaacggaag aaaagctggg tgcggcctgt gaggatcagc ttatcgatga taagctgtca
      601 aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
      661 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
      721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      781 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     1381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1501 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttgaa gctgtccctg atggtcgtca tctacctgcc
     2521 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     2581 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     2641 ccccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     2701 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     2761 ggtgaatccg ttagcgaggt gccgccctgc ttcatccccg tggcccgttg ctcgcgtttg
     2821 ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc
     2881 ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt
     2941 ccgaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc
     3001 agcgacccgc ttaacagcgt caacagcgtg ccgcagatcc cggggggcaa tgagatatga
     3061 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     3121 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     3181 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     3241 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     3301 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     3361 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     3421 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     3481 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     3541 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     3601 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     3661 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     3721 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     3781 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     3841 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     3901 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     3961 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4021 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccggat cgggagatgg
     4081 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4141 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4201 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4261 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4321 caacgtcggg gcggcaggcc ctgccatagc cactggcccc gtgggttagg gacggggtcc
     4381 cccatgggga atggtttatg gttcgtgggg gttattattt tgggcgttgc gtggggtcag
     4441 gtccacgact ggactgagca gacagaccca tggtttttgg atggcctggg catggaccgc
     4501 atgtactggc gcgacacgaa caccgggcgt ctgtggctgc caaacacccc cgacccccaa
     4561 aaaccaccgc gcggatttct ggcgtgccaa gctagtcgac caattctcat gtttgacagc
     4621 ttatcatcgc agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta
     4681 tggaagatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     4741 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     4801 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
     4861 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4921 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     4981 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     5041 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     5101 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     5161 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     5221 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
     5281 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     5341 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
     5401 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tctctagaag ctgggtacca
     5461 gctgctagcg ccaccatgga gaccaccccc ctgaacagtc aaaagcaact ctctgcctgt
     5521 gaggatggag aggactgtca ggagaatggc gttcttcaaa aggttgtacc cacccctgga
     5581 gataaggtgg aaagcggcca gatttccaac ggctacagcg ccgtgccatc acctggagcc
     5641 ggggatgata cacggcacag cattcccgca accaccacga cactggttgc agagcttcac
     5701 cagggcgaga gggagacgtg gggcaaaaag gtagattttc tgctttccgt gatcggttac
     5761 gcggtcgatc tggggaacgt gtggaggttc ccttacattt gttaccaaaa cggagggggt
     5821 gccttcttgc tgccctacac aattatggcg atttttgggg gtatcccgct cttctacatg
     5881 gaactggcct tgggtcagta ccaccggaac ggctgcatta gcatttggcg caagatatgc
     5941 cccattttca agggaatcgg atacgcaatc tgtataatcg cgttctatat tgcatcatat
     6001 tacaacacca tcatggcctg ggccctttat tacttgatca gcagtttcac cgaccaattg
     6061 ccgtggacga gctgtaagaa ctcatggaat accgggaact gtaccaacta cttctctgag
     6121 gacagctcag gaggtagcgg gggtgaacag aagctgatta gcgaggagga cttgggcagt
     6181 ccaggcggaa gcggcaacat cacttggaca ctccatagta cgagccccgc agaggagttt
     6241 tacaccaggc acgtactgca gattcatagg agcaagggtc tgcaagacct gggcggtatt
     6301 agctggcagc tcgcattgtg cataatgttg atctttacgg tgatctattt ctctatctgg
     6361 aagggagtaa agacatccgg caaggtggtg tgggtgacgg cgaccttccc ctacattata
     6421 ctcagcgtgt tgttggtgag gggggcaacc ctgcctgggg cctggcgagg cgtgctgttc
     6481 tatctcaagc caaactggca gaagcttctt gaaaccggag tgtggatcga cgcggccgcg
     6541 caaatcttct tttccctcgg tcccggcttc ggggtccttc tggcattcgc aagctacaac
     6601 aagttcaaca ataactgcta ccaggacgcc ctcgtgacaa gtgtagtgaa ctgcatgacc
     6661 tctttcgtga gcgggtttgt gatcttcaca gtactcggat atatggctga gatgaggaac
     6721 gaagatgtta gtgaggttgc caaagacgct ggcccgagcc tgttgttcat cacatacgcc
     6781 gaggctatcg cgaacatgcc cgcatctact ttcttcgcca tcatcttttt cctgatgctg
     6841 ataacccttg ggctggatag caccttcgcc ggactggagg gtgtcatcac tgccgttctc
     6901 gacgagttcc cccatgtgtg ggccaagcga cgggaaaggt tcgtgctggc cgtcgtaatc
     6961 acacacttct tcggctcact ggtgaccctt acctttggcg gtgcgtacgt ggtcaagctg
     7021 ctcgaagagt acgccactgg accggcggtt ctgaccgtgg cattgatcga ggccgtggcc
     7081 gtaagctggt tttacggcat cacgcagttc tgtagggacg tgaaagagat gctgggcttt
     7141 tctcccggtt ggttttggag gatttgttgg gtagcaatta gccccctgtt tctccttttc
     7201 attatttgct cttttctgat gtctccaccg cagctccgac tgtttcagta caactacccc
     7261 tactggtcaa ttatcctggg ctattgtatc ggcacctcaa gtttcatctg catccccaca
     7321 tacatcgcat acagactgat aattacgccc ggtactttta aggagcgaat cattaaaagc
     7381 ataacccccg aaacccccac cgagatccct tgtggcgata taagactcaa tgcggtctaa
     7441 tagctcgagg ccggcaaggc cggatccaga catgataaga tacattgatg agtttggaca
     7501 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     7561 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     7621 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     7681 atgtggtatg gctgattatg atccggctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     7741 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     7801 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgagg
     7861 tcgactctag aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc
     7921 cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg
     7981 ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag
     8041 ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag
     8101 cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga
     8161 agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag
     8221 gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg
     8281 gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc
     8341 ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg
     8401 ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc
     8461 caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa
     8521 ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa
     8581 tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa
     8641 cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa
     8701 tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc
     8761 ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa
     8821 tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta
     8881 tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca
     8941 gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg
     9001 gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc
     9061 tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg
     9121 agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg
     9181 tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa
     9241 aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg
     9301 cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag
     9361 ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg
     9421 gtagcatata ctacccaaat atctggatag catatgctat cctaatctat atctgggtag
     9481 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     9541 tatatgctat cctaatttat atctgggtag cataggctat cctaatct
//
