LOCUS       sequence-205587-        9685 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for codon optimized mutant hSERT with 
            internal MYC tag inserted in ECL2, mutation R464Q approximately 
            wildtype transport ability.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9685)
  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 9685)
  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-SERTR464Q-myc
FEATURES             Location/Qualifiers
     source          1..9685
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(757..775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        843..947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(948..1016,1017..1808)
                     /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(1166..1185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1979..2567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2468..2487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2998..3143
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3186..4223
                     /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(4221..4240)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4265..4313
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4967..5346
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5347..5550
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5501..5521
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5547..5571
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5599..5608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6274..6303
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    7592..7726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7616..7635)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7670..7689
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7869..7891)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctcctcctgc tcctgcccct cctgctcctg cccctcccgc tcctgctcct gctcctgttc
       61 caccgtgggt ccctttgcag ccaatgcaac ttggacgttt ttggggtctc cggacaccat
      121 ctctatgtct tggccctgat cctgagccgc ccggggctcc tggtcttccg cctcctcgtc
      181 ctcgtcctct tccccgtcct cgtccatggt tatcaccccc tcttctttga ggtccactgc
      241 cgccggagcc ttctggtcca gatgtgtctc ccttctctcc taggccattt ccaggtcctg
      301 tacctggccc ctcgtcagac atgattcaca ctaaaagaga tcaatagaca tctttattag
      361 acgacgctca gtgaatacag ggagtgcaga ctcctgcccc ctccaacagc ccccccaccc
      421 tcatcccctt catggtcgct gtcagacaga tccaggtctg aaaattcccc atcctccgaa
      481 ccatcctcgt cctcatcacc aattactcgc agcccggaaa actcccgctg aacatcctca
      541 agatttgcgt cctgagcctc aagccaggcc tcaaattcct cgtccccctt tttgctggac
      601 ggtagggatg gggattctcg ggacccctcc tcttcctctt caaggtcacc agacagagat
      661 gctactgggg caacggaaga aaagctgggt gcggcctgtg aggatcagct tatcgatgat
      721 aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
      781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     1501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2101 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2581 cggccttttt acggttcctg gccttttgct ggccttgaag ctgtccctga tggtcgtcat
     2641 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     2701 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     2761 gcgcgtcggc cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     2821 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     2881 tcttggagtg gtgaatccgt tagcgaggtg ccgccctgct tcatccccgt ggcccgttgc
     2941 tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg
     3001 acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg
     3061 cggcgcggtc cgaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag
     3121 cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc ggggggcaat
     3181 gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     3241 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     3301 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     3361 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     3421 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     3481 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     3541 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     3601 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     3661 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     3721 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     3781 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     3841 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     3901 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     3961 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     4021 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     4081 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     4141 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccggatc
     4201 gggagatggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4261 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     4321 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4381 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4441 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc actggccccg tgggttaggg
     4501 acggggtccc ccatggggaa tggtttatgg ttcgtggggg ttattatttt gggcgttgcg
     4561 tggggtcagg tccacgactg gactgagcag acagacccat ggtttttgga tggcctgggc
     4621 atggaccgca tgtactggcg cgacacgaac accgggcgtc tgtggctgcc aaacaccccc
     4681 gacccccaaa aaccaccgcg cggatttctg gcgtgccaag ctagtcgacc aattctcatg
     4741 tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac
     4801 tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat attagtcatt
     4861 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     4921 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     4981 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     5041 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     5101 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     5161 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     5221 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     5281 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     5341 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     5401 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     5461 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     5521 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat ctctagaagc
     5581 tgggtaccag ctgctagcgc caccatggag accacccccc tgaacagtca aaagcaactc
     5641 tctgcctgtg aggatggaga ggactgtcag gagaatggcg ttcttcaaaa ggttgtaccc
     5701 acccctggag ataaggtgga aagcggccag atttccaacg gctacagcgc cgtgccatca
     5761 cctggagccg gggatgatac acggcacagc attcccgcaa ccaccacgac actggttgca
     5821 gagcttcacc agggcgagag ggagacgtgg ggcaaaaagg tagattttct gctttccgtg
     5881 atcggttacg cggtcgatct ggggaacgtg tggaggttcc cttacatttg ttaccaaaac
     5941 ggagggggtg ccttcttgct gccctacaca attatggcga tttttggggg tatcccgctc
     6001 ttctacatgg aactggcctt gggtcagtac caccggaacg gctgcattag catttggcgc
     6061 aagatatgcc ccattttcaa gggaatcgga tacgcaatct gtataatcgc gttctatatt
     6121 gcatcatatt acaacaccat catggcctgg gccctttatt acttgatcag cagtttcacc
     6181 gaccaattgc cgtggacgag ctgtaagaac tcatggaata ccgggaactg taccaactac
     6241 ttctctgagg acagctcagg aggtagcggg ggtgaacaga agctgattag cgaggaggac
     6301 ttgggcagtc caggcggaag cggcaacatc acttggacac tccatagtac gagccccgca
     6361 gaggagtttt acaccaggca cgtactgcag attcatagga gcaagggtct gcaagacctg
     6421 ggcggtatta gctggcagct cgcattgtgc ataatgttga tctttacggt gatctatttc
     6481 tctatctgga agggagtaaa gacatccggc aaggtggtgt gggtgacggc gaccttcccc
     6541 tacattatac tcagcgtgtt gttggtgagg ggggcaaccc tgcctggggc ctggcgaggc
     6601 gtgctgttct atctcaagcc aaactggcag aagcttcttg aaaccggagt gtggatcgac
     6661 gcggccgcgc aaatcttctt ttccctcggt cccggcttcg gggtccttct ggcattcgca
     6721 agctacaaca agttcaacaa taactgctac caggacgccc tcgtgacaag tgtagtgaac
     6781 tgcatgacct ctttcgtgag cgggtttgtg atcttcacag tactcggata tatggctgag
     6841 atgaggaacg aagatgttag tgaggttgcc aaagacgctg gcccgagcct gttgttcatc
     6901 acatacgccg aggctatcgc gaacatgccc gcatctactt tcttcgccat catctttttc
     6961 ctgatgctga taacccttgg gctggatagc accttcgccg gactggaggg tgtcatcact
     7021 gccgttctcg acgagttccc ccatgtgtgg gccaagcgac gggaacagtt cgtgctggcc
     7081 gtcgtaatca catgcttctt cggctcactg gtgaccctta cctttggcgg tgcgtacgtg
     7141 gtcaagctgc tcgaagagta cgccactgga ccggcggttc tgaccgtggc attgatcgag
     7201 gccgtggccg taagctggtt ttacggcatc acgcagttct gtagggacgt gaaagagatg
     7261 ctgggctttt ctcccggttg gttttggagg atttgttggg tagcaattag ccccctgttt
     7321 ctccttttca ttatttgctc ttttctgatg tctccaccgc agctccgact gtttcagtac
     7381 aactacccct actggtcaat tatcctgggc tattgtatcg gcacctcaag tttcatctgc
     7441 atccccacat acatcgcata cagactgata attacgcccg gtacttttaa ggagcgaatc
     7501 attaaaagca taacccccga aacccccacc gagatccctt gtggcgatat aagactcaat
     7561 gcggtctaat agctcgaggc cggcaaggcc ggatccagac atgataagat acattgatga
     7621 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     7681 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     7741 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     7801 cctctacaaa tgtggtatgg ctgattatga tccggctgcc tcgcgcgttt cggtgatgac
     7861 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     7921 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     7981 gccatgaggt cgactctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
     8041 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
     8101 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
     8161 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
     8221 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
     8281 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
     8341 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
     8401 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
     8461 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
     8521 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
     8581 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
     8641 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
     8701 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
     8761 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
     8821 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
     8881 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
     8941 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
     9001 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
     9061 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
     9121 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
     9181 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
     9241 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
     9301 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
     9361 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
     9421 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
     9481 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
     9541 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     9601 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     9661 tctgggtagt atatgctatc ctaat
//
