LOCUS       sequence-205616-        9662 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for codon optimized mutant hSERT with 
            internal MYC tag inserted in ECL2, mutation I576V approximately 
            wildtype transport.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9662)
  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 9662)
  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-SERTI576V-myc
FEATURES             Location/Qualifiers
     source          1..9662
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(719..737)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        805..909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(910..978,979..1770)
                     /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(1128..1147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1941..2529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2430..2449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2960..3105
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3148..4185
                     /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(4183..4202)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4227..4275
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4929..5308
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5309..5512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5463..5483
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5509..5533
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5561..5570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6236..6265
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    7554..7688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7578..7597)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7632..7651
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7831..7853)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gctcctgctc ctgctcctgt tccaccgtgg gtccctttgc agccaatgca acttggacgt
       61 ttttggggtc tccggacacc atctctatgt cttggccctg atcctgagcc gcccggggct
      121 cctggtcttc cgcctcctcg tcctcgtcct cttccccgtc ctcgtccatg gttatcaccc
      181 cctcttcttt gaggtccact gccgccggag ccttctggtc cagatgtgtc tcccttctct
      241 cctaggccat ttccaggtcc tgtacctggc ccctcgtcag acatgattca cactaaaaga
      301 gatcaataga catctttatt agacgacgct cagtgaatac agggagtgca gactcctgcc
      361 ccctccaaca gcccccccac cctcatcccc ttcatggtcg ctgtcagaca gatccaggtc
      421 tgaaaattcc ccatcctccg aaccatcctc gtcctcatca ccaattactc gcagcccgga
      481 aaactcccgc tgaacatcct caagatttgc gtcctgagcc tcaagccagg cctcaaattc
      541 ctcgtccccc tttttgctgg acggtaggga tggggattct cgggacccct cctcttcctc
      601 ttcaaggtca ccagacagag atgctactgg ggcaacggaa gaaaagctgg gtgcggcctg
      661 tgaggatcag cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc
      721 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      781 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      841 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      901 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      961 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1021 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1081 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1141 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1201 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1261 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1321 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1381 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1441 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1501 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1561 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1621 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1681 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1741 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1801 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1861 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1921 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1981 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2041 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2101 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2161 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2221 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2281 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2341 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2401 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2461 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2521 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggccttga
     2581 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     2641 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     2701 aacgccagca agacgtagcc cagcgcgtcg gccccgagat gcgccgcgtg cggctgctgg
     2761 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     2821 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccctg
     2881 cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt
     2941 gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc
     3001 gaacacgcag atgcagtcgg ggcggcgcgg tccgaggtcc acttcgcata ttaaggtgac
     3061 gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg tcaacagcgt
     3121 gccgcagatc ccggggggca atgagatatg aaaaagcctg aactcaccgc gacgtctgtc
     3181 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     3241 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     3301 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     3361 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     3421 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     3481 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     3541 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     3601 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     3661 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     3721 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     3781 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     3841 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     3901 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     3961 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     4021 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     4081 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     4141 cgccccagca ctcgtccgga tcgggagatg ggggaggcta actgaaacac ggaaggagac
     4201 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     4261 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4321 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4381 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4441 ccactggccc cgtgggttag ggacggggtc ccccatgggg aatggtttat ggttcgtggg
     4501 ggttattatt ttgggcgttg cgtggggtca ggtccacgac tggactgagc agacagaccc
     4561 atggtttttg gatggcctgg gcatggaccg catgtactgg cgcgacacga acaccgggcg
     4621 tctgtggctg ccaaacaccc ccgaccccca aaaaccaccg cgcggatttc tggcgtgcca
     4681 agctagtcga ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt
     4741 gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga atcaatattg
     4801 gcaattagcc atattagtca ttggttatat agcataaatc aatattggct attggccatt
     4861 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
     4921 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     4981 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5041 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5101 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5161 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     5221 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     5281 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     5341 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     5401 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     5461 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     5521 gaaccgtcag atctctagaa gctgggtacc agctgctagc gccaccatgg agaccacccc
     5581 cctgaacagt caaaagcaac tctctgcctg tgaggatgga gaggactgtc aggagaatgg
     5641 cgttcttcaa aaggttgtac ccacccctgg agataaggtg gaaagcggcc agatttccaa
     5701 cggctacagc gccgtgccat cacctggagc cggggatgat acacggcaca gcattcccgc
     5761 aaccaccacg acactggttg cagagcttca ccagggcgag agggagacgt ggggcaaaaa
     5821 ggtagatttt ctgctttccg tgatcggtta cgcggtcgat ctggggaacg tgtggaggtt
     5881 cccttacatt tgttaccaaa acggaggggg tgccttcttg ctgccctaca caattatggc
     5941 gatttttggg ggtatcccgc tcttctacat ggaactggcc ttgggtcagt accaccggaa
     6001 cggctgcatt agcatttggc gcaagatatg ccccattttc aagggaatcg gatacgcaat
     6061 ctgtataatc gcgttctata ttgcatcata ttacaacacc atcatggcct gggcccttta
     6121 ttacttgatc agcagtttca ccgaccaatt gccgtggacg agctgtaaga actcatggaa
     6181 taccgggaac tgtaccaact acttctctga ggacagctca ggaggtagcg ggggtgaaca
     6241 gaagctgatt agcgaggagg acttgggcag tccaggcgga agcggcaaca tcacttggac
     6301 actccatagt acgagccccg cagaggagtt ttacaccagg cacgtactgc agattcatag
     6361 gagcaagggt ctgcaagacc tgggcggtat tagctggcag ctcgcattgt gcataatgtt
     6421 gatctttacg gtgatctatt tctctatctg gaagggagta aagacatccg gcaaggtggt
     6481 gtgggtgacg gcgaccttcc cctacattat actcagcgtg ttgttggtga ggggggcaac
     6541 cctgcctggg gcctggcgag gcgtgctgtt ctatctcaag ccaaactggc agaagcttct
     6601 tgaaaccgga gtgtggatcg acgcggccgc gcaaatcttc ttttccctcg gtcccggctt
     6661 cggggtcctt ctggcattcg caagctacaa caagttcaac aataactgct accaggacgc
     6721 cctcgtgaca agtgtagtga actgcatgac ctctttcgtg agcgggtttg tgatcttcac
     6781 agtactcgga tatatggctg agatgaggaa cgaagatgtt agtgaggttg ccaaagacgc
     6841 tggcccgagc ctgttgttca tcacatacgc cgaggctatc gcgaacatgc ccgcatctac
     6901 tttcttcgcc atcatctttt tcctgatgct gataaccctt gggctggata gcaccttcgc
     6961 cggactggag ggtgtcatca ctgccgttct cgacgagttc ccccatgtgt gggccaagcg
     7021 acgggaaagg ttcgtgctgg ccgtcgtaat cacatgcttc ttcggctcac tggtgaccct
     7081 tacctttggc ggtgcgtacg tggtcaagct gctcgaagag tacgccactg gaccggcggt
     7141 tctgaccgtg gcattgatcg aggccgtggc cgtaagctgg ttttacggca tcacgcagtt
     7201 ctgtagggac gtgaaagaga tgctgggctt ttctcccggt tggttttgga ggatttgttg
     7261 ggtagcaatt agccccctgt ttctcctttt cattatttgc tcttttctga tgtctccacc
     7321 gcagctccga ctgtttcagt acaactaccc ctactggtca attgtcctgg gctattgtat
     7381 cggcacctca agtttcatct gcatccccac atacatcgca tacagactga taattacgcc
     7441 cggtactttt aaggagcgaa tcattaaaag cataaccccc gaaaccccca ccgagatccc
     7501 ttgtggcgat ataagactca atgcggtcta atagctcgag gccggcaagg ccggatccag
     7561 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     7621 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     7681 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     7741 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatccggctg
     7801 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     7861 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     7921 tgttggcggg tgtcggggcg cagccatgag gtcgactcta gaggatcgat ccccgccccg
     7981 gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc
     8041 ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg
     8101 gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat
     8161 ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac
     8221 aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt
     8281 acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg
     8341 tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc
     8401 ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta
     8461 accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag
     8521 taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta
     8581 catggggtca ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa
     8641 gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt
     8701 ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa
     8761 caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg
     8821 tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat
     8881 gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac
     8941 tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg
     9001 atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta
     9061 cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt
     9121 gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga
     9181 caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc
     9241 cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc
     9301 taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct
     9361 gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt
     9421 acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg
     9481 ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata
     9541 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     9601 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     9661 gc
//
