LOCUS       sequence-124851-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of hSERT with C-terminal YFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Just H, Sitte HH, Schmid JA, Freissmuth M, Kudlacek O
  TITLE     Identification of an additional interaction domain in transmembrane 
            domains 11 and 12 that supports oligomer formation in the human 
            serotonin transporter.
  JOURNAL   J Biol Chem. 2004 Feb 20;279(8):6650-7. Epub 2003 Dec 5.
  PUBMED    14660642
REFERENCE   2  (bases 1 to 6640)
  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: "J Biol 
            Chem."; date: "2004-02-20"; volume: "279(8)"; pages: "6650-7. Epub 
            2003 Dec 5"
COMMENT     Sequence Label: hSERT-N1-pEYFP
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2580..2589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2586..2588,2589..2591,2592..3305)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2631..2652)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2892..2911)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3239..3260
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3428..3549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3465..3484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3519..3538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3556..4011)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3693..3714)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3905..3924
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4038..4142
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4144..4501
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4144..4335
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4167..4187)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4352..4487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4536..5330
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4536..5330
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4590..4609)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5200..5219
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5518..5537)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5562..5609
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5938..6526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6427..6446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcta caccatccgt cccgatggag acgacgccct
      661 tgaattctca gaagcagcta tcagcgtgtg aagatggaga agattgtcag gaaaacggag
      721 ttctacagaa ggttgttccc accccagggg acaaagtgga gtccgggcaa atatccaatg
      781 ggtactcagc agttccaagt cctggtgcgg gagatgacac acggcactct atcccagcga
      841 ccaccaccac cctagtggct gagcttcatc aaggggaacg ggagacctgg ggcaagaagg
      901 tggatttcct tctctcagtg attggctatg ctgtggacct gggcaatgtc tggcgcttcc
      961 cctacatatg ttaccagaat ggaggggggg cattcctcct cccctacacc atcatggcca
     1021 tttttggggg aatcccgctc ttttacatgg agctcgcact gggacagtac caccgaaatg
     1081 gatgcatttc aatatggagg aaaatctgcc cgattttcaa agggattggt tatgccatct
     1141 gcatcattgc cttttacatt gcttcctact acaacaccat catggcctgg gcgctatact
     1201 acctcatctc ctccttcacg gaccagctgc cctggaccag ctgcaagaac tcctggaaca
     1261 ctggcaactg caccaattac ttctccgagg acaacatcac ctggaccctc cattccacgt
     1321 cccctgctga agaattttac acgcgccacg tcctgcagat ccaccggtct aaggggctcc
     1381 aggacctggg gggcatcagc tggcagctgg ccctctgcat catgctgatc ttcactgtta
     1441 tctacttcag catctggaaa ggcgtcaaga cctctggcaa ggtggtgtgg gtgacagcca
     1501 ccttccctta tatcatcctt tctgtcctgc tggtgagggg tgccaccctc cctggagcct
     1561 ggaggggtgt tctcttctac ttgaaaccca attggcagaa actcctggag acaggggtgt
     1621 ggatagatgc agccgctcag atcttcttct ctcttggtcc gggctttggg gtcctgctgg
     1681 cttttgctag ctacaacaag ttcaacaaca actgctacca agatgccctg gtgaccagcg
     1741 tggtgaactg catgacgagc ttcgtttcgg gatttgtcat cttcacagtg ctcggttaca
     1801 tggctgagat gaggaatgaa gatgtgtctg aggtggccaa agacgcaggt cccagcctcc
     1861 tcttcatcac gtatgcagaa gcgatagcca acatgccagc gtccactttc tttgccatca
     1921 tcttctttct gatgttaatc acgctgggct tggacagcac gtttgcaggc ttggaggggg
     1981 tgatcacggc tgtgctggat gagttcccac acgtctgggc caagcgccgg gagcggttcg
     2041 tgctcgccgt ggtcatcacc tgcttctttg gatccctggt caccctgact tttggagggg
     2101 cctacgtggt gaagctgctg gaggagtatg ccacggggcc cgcagtgctc actgtcgcgc
     2161 tgatcgaagc agtcgctgtg tcttggttct atggcatcac tcagttctgc agggacgtga
     2221 aggaaatgct cggcttcagc ccggggtggt tctggaggat ctgctgggtg gccatcagcc
     2281 ctctgtttct cctgttcatc atttgcagtt ttctgatgag cccgccacaa ctacgacttt
     2341 tccaatataa ttatccttac tggagtatca tcttgggtta ctgcatagga acctcatctt
     2401 tcatttgcat ccccacatat atagcttatc ggttgatcat cactccaggg acatttaaag
     2461 agcgtattat taaaagtatt accccagaaa caccaacaga aattccttgt ggggacatcc
     2521 gcttgaatgc tgtgggaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg
     2581 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2641 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2701 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2761 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     2821 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2881 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2941 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3001 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3061 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3121 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3181 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3241 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3301 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3361 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3421 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3481 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3541 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3661 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3721 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3781 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3841 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3901 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3961 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4141 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4321 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4441 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4501 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4561 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4621 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4681 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4741 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4801 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4861 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4921 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4981 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5041 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5101 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5161 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5221 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5281 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5341 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5401 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5461 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5521 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5581 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5641 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5701 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5761 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5821 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5881 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5941 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6001 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6061 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6121 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6181 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6241 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6301 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6361 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6421 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6481 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6541 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6601 atcccctgat tctgtggata accgtattac cgccatgcat
//
