LOCUS       sequence-127692-        6623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of hSERT with N-terminal YFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6623)
  AUTHORS   Sucic S, Dallinger S, Zdrazil B, Weissensteiner R, Jorgensen TN, 
            Holy M, Kudlacek O, Seidel S, Cha JH, Gether U, Newman AH, Ecker GF,
            Freissmuth M, Sitte HH
  TITLE     The N terminus of monoamine transporters is a lever required for the
            action of amphetamines.
  JOURNAL   J Biol Chem. 2010 Apr 2;285(14):10924-38. doi: 
            10.1074/jbc.M109.083154. Epub 2010 Jan 29.
  PUBMED    20118234
REFERENCE   2  (bases 1 to 6623)
  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"; doi: 
            "10.1074/jbc.M109.083154"; journalName: "J Biol Chem"; date: 
            "2010-04-2- 2"; volume: "285"; issue: "14"; pages: "10924-38"
COMMENT     Sequence Label: pEYFP-C1-hSERT
FEATURES             Location/Qualifiers
     source          1..6623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(243..262)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    287..334
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      663..1251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1152..1171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1426..1729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1730..1933
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1884..1904
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1972..1981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1978..1980,1981..1983,1984..2694)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2023..2044)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2284..2303)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2631..2652
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4776..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4813..4832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4867..4886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4904..5359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5041..5062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5253..5272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5386..5490
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5492..5849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5492..5683
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5515..5535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5700..5835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5884..6623,1..55)
                     /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             join(5884..6623,1..55)
                     /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(5938..5957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6548..6567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
       61 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
      121 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
      181 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
      241 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
      301 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
      361 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
      421 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
      481 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
      541 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      781 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
      841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1321 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     1381 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1441 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1501 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1561 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1621 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1681 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1741 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1801 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1861 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1921 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
     1981 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2041 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2101 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2161 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     2221 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2281 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2341 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2401 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2461 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2521 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2581 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2641 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     2701 ctcagatctc gagccatgga gacgacgccc ttgaattctc agaagcagct atcagcgtgt
     2761 gaagatggag aagattgtca ggaaaacgga gttctacaga aggttgttcc caccccaggg
     2821 gacaaagtgg agtccgggca aatatccaat gggtactcag cagttccaag tcctggtgcg
     2881 ggagatgaca cacggcactc tatcccagcg accaccacca ccctagtggc tgagcttcat
     2941 caaggggaac gggagacctg gggcaagaag gtggatttcc ttctctcagt gattggctat
     3001 gctgtggacc tgggcaatgt ctggcgcttc ccctacatat gttaccagaa tggagggggg
     3061 gcattcctcc tcccctacac catcatggcc atttttgggg gaatcccgct cttttacatg
     3121 gagctcgcac tgggacagta ccaccgaaat ggatgcattt caatatggag gaaaatctgc
     3181 ccgattttca aagggattgg ttatgccatc tgcatcattg ccttttacat tgcttcctac
     3241 tacaacacca tcatggcctg ggcgctatac tacctcatct cctccttcac ggaccagctg
     3301 ccctggacca gctgcaagaa ctcctggaac actggcaact gcaccaatta cttctccgag
     3361 gacaacatca cctggaccct ccattccacg tcccctgctg aagaatttta cacgcgccac
     3421 gtcctgcaga tccaccggtc taaggggctc caggacctgg ggggcatcag ctggcagctg
     3481 gccctctgca tcatgctgat cttcactgtt atctacttca gcatctggaa aggcgtcaag
     3541 acctctggca aggtggtgtg ggtgacagcc accttccctt atatcatcct ttctgtcctg
     3601 ctggtgaggg gtgccaccct ccctggagcc tggaggggtg ttctcttcta cttgaaaccc
     3661 aattggcaga aactcctgga gacaggggtg tggatagatg cagccgctca gatcttcttc
     3721 tctcttggtc cgggctttgg ggtcctgctg gcttttgcta gctacaacaa gttcaacaac
     3781 aactgctacc aagatgccct ggtgaccagc gtggtgaact gcatgacgag cttcgtttcg
     3841 ggatttgtca tcttcacagt gctcggttac atggctgaga tgaggaatga agatgtgtct
     3901 gaggtggcca aagacgcagg tcccagcctc ctcttcatca cgtatgcaga agcgatagcc
     3961 aacatgccag cgtccacttt ctttgccatc atcttctttc tgatgttaat cacgctgggc
     4021 ttggacagca cgtttgcagg cttggagggg gtgatcacgg ctgtgctgga tgagttccca
     4081 cacgtctggg ccaagcgccg ggagcggttc gtgctcgccg tggtcatcac ctgcttcttt
     4141 ggatccctgg tcaccctgac ttttggaggg gcctacgtgg tgaagctgct ggaggagtat
     4201 gccacggggc ccgcagtgct cactgtcgcg ctgatcgaag cagtcgctgt gtcttggttc
     4261 tatggcatca ctcagttctg cagggacgtg aaggaaatgc tcggcttcag cccggggtgg
     4321 ttctggagga tctgctgggt ggccatcagc cctctgtttc tcctgttcat catttgcagt
     4381 tttctgatga gcccgccaca actacgactt ttccaatata attatcctta ctggagtatc
     4441 atcttgggtt actgcatagg aacctcatct ttcatttgca tccccacata tatagcttat
     4501 cggttgatca tcactccagg gacatttaaa gagcgtatta ttaaaagtat taccccagaa
     4561 acaccaacag aaattccttg tggggacatc cgcttgaatg ctgtgtaaaa gcttcgaatt
     4621 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     4681 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4741 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5341 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5401 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5461 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5821 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5881 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5941 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6001 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6061 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6121 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6181 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6241 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6301 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6361 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     6421 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6481 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6541 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6601 ttcctcgtgc tttacggtat cgc
//
