LOCUS       sequence-127668-        6623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of hSERT with N-terminal CFP 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: pECFP-C1-hSERT
FEATURES             Location/Qualifiers
     source          1..6623
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3411..3532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3448..3467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3502..3521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3539..3994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3676..3697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3888..3907
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4021..4125
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4127..4484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4127..4318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4150..4170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4335..4470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4519..5313
                     /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             4519..5313
                     /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(4573..4592)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5183..5202
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5501..5520)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5545..5592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5921..6509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6410..6429
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagcc atggagacga cgcccttgaa ttctcagaag
     1381 cagctatcag cgtgtgaaga tggagaagat tgtcaggaaa acggagttct acagaaggtt
     1441 gttcccaccc caggggacaa agtggagtcc gggcaaatat ccaatgggta ctcagcagtt
     1501 ccaagtcctg gtgcgggaga tgacacacgg cactctatcc cagcgaccac caccacccta
     1561 gtggctgagc ttcatcaagg ggaacgggag acctggggca agaaggtgga tttccttctc
     1621 tcagtgattg gctatgctgt ggacctgggc aatgtctggc gcttccccta catatgttac
     1681 cagaatggag ggggggcatt cctcctcccc tacaccatca tggccatttt tgggggaatc
     1741 ccgctctttt acatggagct cgcactggga cagtaccacc gaaatggatg catttcaata
     1801 tggaggaaaa tctgcccgat tttcaaaggg attggttatg ccatctgcat cattgccttt
     1861 tacattgctt cctactacaa caccatcatg gcctgggcgc tatactacct catctcctcc
     1921 ttcacggacc agctgccctg gaccagctgc aagaactcct ggaacactgg caactgcacc
     1981 aattacttct ccgaggacaa catcacctgg accctccatt ccacgtcccc tgctgaagaa
     2041 ttttacacgc gccacgtcct gcagatccac cggtctaagg ggctccagga cctggggggc
     2101 atcagctggc agctggccct ctgcatcatg ctgatcttca ctgttatcta cttcagcatc
     2161 tggaaaggcg tcaagacctc tggcaaggtg gtgtgggtga cagccacctt cccttatatc
     2221 atcctttctg tcctgctggt gaggggtgcc accctccctg gagcctggag gggtgttctc
     2281 ttctacttga aacccaattg gcagaaactc ctggagacag gggtgtggat agatgcagcc
     2341 gctcagatct tcttctctct tggtccgggc tttggggtcc tgctggcttt tgctagctac
     2401 aacaagttca acaacaactg ctaccaagat gccctggtga ccagcgtggt gaactgcatg
     2461 acgagcttcg tttcgggatt tgtcatcttc acagtgctcg gttacatggc tgagatgagg
     2521 aatgaagatg tgtctgaggt ggccaaagac gcaggtccca gcctcctctt catcacgtat
     2581 gcagaagcga tagccaacat gccagcgtcc actttctttg ccatcatctt ctttctgatg
     2641 ttaatcacgc tgggcttgga cagcacgttt gcaggcttgg agggggtgat cacggctgtg
     2701 ctggatgagt tcccacacgt ctgggccaag cgccgggagc ggttcgtgct cgccgtggtc
     2761 atcacctgct tctttggatc cctggtcacc ctgacttttg gaggggccta cgtggtgaag
     2821 ctgctggagg agtatgccac ggggcccgca gtgctcactg tcgcgctgat cgaagcagtc
     2881 gctgtgtctt ggttctatgg catcactcag ttctgcaggg acgtgaagga aatgctcggc
     2941 ttcagcccgg ggtggttctg gaggatctgc tgggtggcca tcagccctct gtttctcctg
     3001 ttcatcattt gcagttttct gatgagcccg ccacaactac gacttttcca atataattat
     3061 ccttactgga gtatcatctt gggttactgc ataggaacct catctttcat ttgcatcccc
     3121 acatatatag cttatcggtt gatcatcact ccagggacat ttaaagagcg tattattaaa
     3181 agtattaccc cagaaacacc aacagaaatt ccttgtgggg acatccgctt gaatgctgtg
     3241 taaaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat
     3301 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3361 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3421 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3481 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3541 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3601 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3661 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3721 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3781 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3841 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3901 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3961 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4021 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4081 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4141 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4201 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4261 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4321 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4381 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4441 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4501 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4561 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4621 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4681 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4741 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4801 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4861 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4921 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4981 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5041 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5101 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5161 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5221 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5281 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5341 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5401 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5461 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5521 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5581 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5641 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5701 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5761 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5821 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5881 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6001 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6061 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6121 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6181 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6241 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6301 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6361 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6421 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6481 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6541 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6601 ataaccgtat taccgccatg cat
//
