LOCUS       sequence-71101-e        8376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-tagged human DAT with first 5 N-terminal serines 
            mutated to aspartates in pCIHygro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8376)
  AUTHORS   Fog JU, Khoshbouei H, Holy M, Owens WA, Vaegter CB, Sen N, 
            Nikandrova Y, Bowton E, McMahon DG, Colbran RJ, Daws LC, Sitte HH, 
            Javitch JA, Galli A, Gether U
  TITLE     Calmodulin kinase II interacts with the dopamine transporter C 
            terminus to regulate amphetamine-induced reverse transport.
  JOURNAL   Neuron. 2006 Aug 17;51(4):417-29.
  PUBMED    16908408
REFERENCE   2  (bases 1 to 8376)
  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: "Neuron."; 
            date: "2006-08-17"; volume: "51(4)"; pages: "417-29"
COMMENT     Sequence Label: YFP-synDAT-S/D
FEATURES             Location/Qualifiers
     source          1..8376
                     /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"
     regulatory      914..923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(920..922,923..925,926..1636)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(965..986)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1226..1245)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1573..1594
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3619..3848
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3831..3850
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3924..4499,4500..4502,4503..4511)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4091..4108)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(5863..5880)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5869..6093
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6140..6159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6219..6235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6219..6235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6232..6254)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6243..6259
                     /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(6267..6273,6274..6291,6292..6297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6312..6333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6450..6467)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6621..7209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6701..6720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7380..8171,8172..8240))
                     /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     8003..8022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8241..8345)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ccgggattta aatgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      961 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1021 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1081 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg
     1141 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1201 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1261 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1321 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1381 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1441 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1501 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     1561 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1621 ggacgagctg tacaaggtta acggtaccac catggataag gacaaatgcg atgtgggact
     1681 gatggacgat gtggttgccc ccgctaagga gcccaacgcc gtgggcccca aggaggtgga
     1741 actgattctc gtgaaggagc agaacggcgt gcagctcacc tccagcacac tgaccaaccc
     1801 taggcagagc cccgtggagg cccaggaccg ggagacctgg ggcaagaaaa tcgacttcct
     1861 gctcagcgtg atcggcttcg ccgtcgacct ggctaacgtt tggcggttcc cctacctgtg
     1921 ctacaagaac ggcggaggcg ctttcctggt gccctacctc ctctttatgg tgatcgctgg
     1981 catgcccctg ttctacatgg agctggccct cggccagttt aaccgggagg gagccgcagg
     2041 tgtgtggaag atctgcccca tcctcaaagg cgtgggattc accgtgatcc tgatcagcct
     2101 gtacgtaggc ttcttttaca acgtcatcat tgcctgggct ctccactacc tgttctcgag
     2161 cttcaccaca gagctgccct ggatccactg caacaatagc tggaacagcc ccaactgcag
     2221 cgatgcgcac cccggcgaca gctccggaga cagctcaggc ctgaacgaca ccttcggaac
     2281 cacacccgcc gctgaatact tcgaacgggg cgtgctgcac ctccatcaga gccacggcat
     2341 cgatgacctt ggtcctccca ggtggcagct gaccgcctgt ctggtgctcg tcatcgtgct
     2401 gctctacttc agcctgtgga agggcgtgaa gacaagcggc aaggtggtct ggatcaccgc
     2461 cacaatgcca tatgtggtcc tgaccgccct gctcctgcgg ggggtgaccc tgcctggggc
     2521 catcgacggc attcgggcct acctgagcgt ggacttctac cggctgtgcg aggctagcgt
     2581 gtggatcgac gctgccactc aagtgtgctt cagcctgggc gtgggattcg gggtcctgat
     2641 cgccttcagc tcctacaaca agttcaccaa caattgctac cgcgacgcca tcgtcaccac
     2701 aagcatcaac tccctgacta gtttttcgag cggctttgtc gtgttcagct ttctgggata
     2761 catggcccag aagcacagcg tgcccatcgg tgacgtcgcc aaggacggcc ccgggctgat
     2821 cttcattatc taccctgagg ccatcgctac cctgcccctg agctccgcct gggctgtcgt
     2881 gttctttatc atgctgctca ccctcggcat cgacagcgct atgggaggca tggagagcgt
     2941 catcaccggc ctgatcgacg aattccagct cctgcaccgg catcgcgagc tgttcaccct
     3001 gttcatcgtg ctggccacct tccttctgag cctgttctgc gtgaccaacg gcggaatcta
     3061 cgtgttcacc ctcctggacc acttcgctgc cggcacctca atattgttcg gcgtgctgat
     3121 cgaggcgatc ggcgtggcct ggttctacgg agtgggccag ttcagcgacg atatccagca
     3181 gatgaccggt cagcggccca gcctgtactg gcggctgtgc tggaagctcg tctccccctg
     3241 cttcctcctg ttcgtggtcg ttgtgtcaat tgtgaccttc cggcctcccc actacggcgc
     3301 ctacatcttc cccgactggg ccaatgcatt gggctgggtg atcgcaacca gctccatggc
     3361 tatggtgccc atctacgctg cctacaagtt ctgtagcctg cccggaagct tccgggagaa
     3421 gctggcgtac gctatcgccc ccgagaagga ccgggagctg gtggaccgcg gcgaggtgcg
     3481 gcagttcacc ctgcggcact ggctgaaggt gtgaagagag tctagaagcg gccgcgtcga
     3541 cggaattcag tggatccact agtaacggcc gccagtgtgc tggaattaat tcgctgtctg
     3601 cgagggccgg ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa
     3661 gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt
     3721 tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt
     3781 gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct
     3841 ctccacaggt gtccactccc aggtccaact gcaggtcgat cgagcatgca tctagggcgg
     3901 ccggccgcac tagaggaatt ccgcccctct ccctcccccc cccctaacgt tactggccga
     3961 agccgcttgg aataaggccg gtgtgtgttt gtctatatgt gattttccac catattgccg
     4021 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     4081 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     4141 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     4201 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     4261 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     4321 ctctcctcaa gcgtagtcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     4381 ggaatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4441 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc
     4501 ttgccacaac ccgtaccaaa gatggataga tccggaaagc ctgaactcac cgcgacgtct
     4561 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     4621 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4681 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     4741 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     4801 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4861 gttctgcacc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     4921 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     4981 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     5041 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     5101 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     5161 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     5221 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5281 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     5341 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5401 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5461 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5521 gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg aacaagagct
     5581 gatttcgaga acgcctcagc cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa
     5641 cggccttacg cttggtggca cagttctcgt ccacagttcg ctaagctcgc tcggctgggt
     5701 cgcgggaggg ccggtcgcag tgattcaggc ccttctggat tgtgttggtc cccagggcac
     5761 gattgtcatg cccacgcact cgggtgatct gactgatccc gcagattgga gatcgccgcc
     5821 cgtgcctgcc gattgggtgc agatctagag ctcgctgatc agcctcgact gtgccttcta
     5881 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5941 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6001 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     6061 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     6121 gctcgagtgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6181 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6241 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6301 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6361 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6421 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6481 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6541 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6601 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6661 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6721 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6781 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6841 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6901 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6961 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7021 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7081 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7141 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7201 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7261 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7321 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7381 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7441 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7501 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7561 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7621 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7681 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7741 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7801 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7861 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7921 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7981 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8041 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8101 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8161 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8221 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8281 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8341 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
