LOCUS       sequence-379872-        7467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of wild-type rat SERT in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  AUTHORS   Singh I, Seth A, Billesbølle CB, Braz J, Rodriguiz RM, Roy K, Bekele
            B, Craik V, Huang X, Boytsov D, Lak P, O'Donnell H, Sandtner W, Roth
            BL, Basbaum AI, Wetsel WC, Manglik A, Shoichet BK, Rudnick G
  TITLE     Structure-based Discovery of Conformationally Selective Inhibitors 
            of the Serotonin Transporter
  JOURNAL   bioRxiv 2022.06.13.495991
  PUBMED    11592963
REFERENCE   2  (bases 1 to 7467)
  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 
            2022.06.13.495991"
COMMENT     Sequence Label: pcDNA3.1_rSERT_WT
FEATURES             Location/Qualifiers
     source          1..7467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     972..988
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     973..989
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1004..1013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1013..1042
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2945..2962
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2969..2992
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3020..3038)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3021..3038)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3058..3075)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3064..3288
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3334..3762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3421..3440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3631..3652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3771..3791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3776..4105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3956..4091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4018..4037
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4172..4966
                     /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(4226..4245)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4836..4855
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5140..5261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5177..5196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5231..5250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5310..5326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5310..5326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5323..5345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5334..5350
                     /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(5358..5364,5365..5382,5383..5388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5403..5424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5541..5558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5792..5811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6471..7262,7263..7331))
                     /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     7094..7113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7332..7436)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
      961 ctggcggccg ctcgaggtcg acggtatcga taagcttgat atcgccacca tggaacaaaa
     1021 actcatttca gaagaggatc tgaacatgga gaccacaccc ttgaattcac agaaagtgct
     1081 gtcagagtgt aaggacagag aggactgtca agaaaatggt gttctacaga agggtgtccc
     1141 caccacagcg gacagggcag agcctagcca aatatccaat gggtactctg cagtccccag
     1201 cacaagtgca ggggacgaag cttcacactc gatcccagct gccaccacca ccctggtggc
     1261 tgagattcgc caaggggagc gggagacctg gggcaagaag atggatttcc tcctgtccgt
     1321 cattggctat gccgtggacc tgggcaacat ctggcggttt ccttacatat gctaccagaa
     1381 tggcggaggg gccttcctcc tcccttatac catcatggcc attttcgggg ggatcccgct
     1441 cttttacatg gagctcgcac tgggccagta ccaccgaaac gggtgcattt ccatatggag
     1501 gaagatctgc ccgattttca aaggcattgg ttacgccatc tgcatcatcg ccttttacat
     1561 cgcctcctac tacaacacca tcatagcctg ggcgctctac tacctcatct cctccctcac
     1621 ggaccggctg ccctggacca gctgcacgaa ctcctggaac actggcaact gcaccaacta
     1681 cttcgcccag gacaacatca cctggacgct gcattccacg tcccccgctg aggagttcta
     1741 cttgcgccat gtcctgcaga tccaccagtc taagggactc caggacctgg gcaccatcag
     1801 ctggcagctg actctctgca tcgtgctcat cttcaccgta atctacttta gcatctggaa
     1861 aggcgtcaaa acatctggca aggtggtgtg ggtgacagcc accttcccat acattgtcct
     1921 ctctgtcctg ctggtgaggg gggccaccct tcctggagcc tggagagggg tcgtcttcta
     1981 cttgaaaccc aactggcaga aactcttgga gacaggggtg tgggtagatg ccgccgctca
     2041 gatcttcttc tctcttggcc cgggctttgg ggttctcctg gcttttgcta gctacaacaa
     2101 gttcaacaac aactgttacc aagatgccct ggtgaccagt gtggtgaact gcatgacaag
     2161 cttcgtctct ggcttcgtca tcttcacggt gcttggctac atggcggaga tgaggaatga
     2221 agatgtgtca gaggtggcca aagacgcagg ccccagcctc ctcttcatca cgtatgcaga
     2281 ggcaattgcc aacatgccag catccacgtt ctttgccatc atcttcttcc tcatgttaat
     2341 cacgctggga ttggacagca cgttcgcagg cctggaaggt gtgatcacag ctgtgctgga
     2401 tgagttccct cacatctggg ccaagcgcag ggaatggttc gtgctcatcg tggtcatcac
     2461 gtgcgtcttg ggatccctgc tcacactgac gtcaggaggg gcatacgtgg tgactctgct
     2521 ggaggagtat gccacggggc cagcagtgct caccgtggcc ctcatcgagg ccgtcgccgt
     2581 gtcttggttc tatggaatca ctcagttctg cagcgatgtg aaggagatgc tgggcttcag
     2641 cccgggatgg ttttggagga tctgctgggt ggccatcagc cctctgtttc tcctgttcat
     2701 catttgcagt tttctgatga gcccacccca gctacggctt ttccaataca actatcccca
     2761 ctggagtatc gtcttgggct actgcatagg gatgtcgtcc gtcatctgca tccctaccta
     2821 tatcatttat cggctgatca gcactccggg gacacttaag gagcgcatta ttaaaagtat
     2881 cactcctgaa acacccacag aaatcccgtg tggggacatc cgcatgaatg ctgtggcggc
     2941 cgcacaccat caccatcacc atggtaccga ctacaaggac gacgatgaca aggaattcta
     3001 ataatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     3061 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3121 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3181 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3241 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3301 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3361 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3421 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3481 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3541 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3601 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3661 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3721 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3781 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3841 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3901 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3961 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4021 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4081 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4141 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4201 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4261 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4321 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4381 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4441 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4501 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4561 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4621 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4681 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4741 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4801 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4861 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4921 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4981 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5101 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5161 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5221 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5281 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tgacgtc
//
