LOCUS       sequence-335066-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expresssion of His-GFP-FCHo2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  AUTHORS   Prasai B, Haber GJ, Strub MP, Ahn R, Ciemniecki JA, Sochacki KA, 
            Taraska JW
  TITLE     The nanoscale molecular morphology of docked exocytic dense-core 
            vesicles in neuroendocrine cells.
  JOURNAL   Nat Commun. 2021 Jun 25;12(1):3970. doi: 10.1038/s41467-021-24167-9.
  PUBMED    34172739
REFERENCE   2  (bases 1 to 7146)
  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: "Nat 
            Commun."; date: "2021-06-25"; volume: "12(1):3970. doi"; pages: " 
            10.1038/s41467-021-24167-9"
COMMENT     Sequence Label: His-GFP-FCHo2
FEATURES             Location/Qualifiers
     source          1..7146
                     /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             625..642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(652..654,655..1365)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(694..715)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(955..974)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1302..1323
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3934..4055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3971..3990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4025..4044
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4062..4517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4199..4220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4411..4430
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4544..4648
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4650..5007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4650..4841
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4673..4693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4858..4993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5042..5836
                     /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             5042..5836
                     /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(5096..5115)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5706..5725
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6024..6043)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6068..6115
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /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 ccatgggcag cagccatcat catcatcatc acagcagcgg cgtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
     1381 cgagctcgga tgggttcgac ctcgggttcg accaagggtc agctgatgaa gcatggacag
     1441 atatcaacta aagaactagc agattttgta agagaaagag ctacaataga agaggcatac
     1501 tccagatcaa tgacaaaact agcgaaatct gcaagcaatt attcacaact tggaacattt
     1561 gcaccaatgt gggatgtatt caaaacatct acagagaaat tagcaaattg tcatttggat
     1621 cttgttagaa aattacagga attaataaag gaagttcaga agtatggaga ggagcaagtc
     1681 aagtctcata agaagactaa agaagaagtc gcagggactc tggaagcagt ccaagccatt
     1741 cagaatataa ctcaggcact ccagaagtcg aaggaaaatt acactgccaa atgtgtagaa
     1801 caggaacgtc tgaaaaagga aggagctaca caaagagaaa tagaaaaggc agctgttaag
     1861 tctaagaaag ctacagatac ttataaactt tatgtggaaa agtatgcatt aacaaaagca
     1921 gattttgagc agaagatgac agaaacagcc cagaaatttc aagatattga agaaactcac
     1981 ctcattcaca taaaggaaat tatcggatcc ttgtcaaatg ctgtcaagga gatccactta
     2041 cagatcggcc aggttcatga agagtttata aataacatgg ctaatactac aattgaaagt
     2101 ttgattcaga aatttgccga gtcaaaaggc acagggaagg aaagacctgg tcttattgaa
     2161 tttgaagagt gtgaccctgc tagtgcagtt gaaggcataa aaccaaggaa aagaaagact
     2221 tttgctttgc ctggaataat taaaaaggaa aaggacgcag aatctgtgga atgtcctgat
     2281 gcagattcgc ttaatattcc agatgtagat gaagaaggtt tcagtattaa accagaagca
     2341 aatcagaatg ataccaaaga gaaccatttc tactcatcca gtgactctga ctctgaagat
     2401 gaagaaccaa agaggtaccg gatagagatc aagcccgcgc atccaaataa cttgcatcac
     2461 acaatggctt ctttggatga actgaaagta tccataggga atataacatt gtccccagca
     2521 gtatctagac acagtccagt gcagatgaat cggaattcat ctaatgaaga attaacaaaa
     2581 tcaaagccgt cttctctacc cactgagaaa gggaccaatg atttacttgc ttgggatccc
     2641 ctgttcggat cgtcgcttga gtcatcttca gctccactaa cttcgtcatc atcggccaga
     2701 cccacgacac ctctctcttt aggcaccctt gttccacctc caaggcctgc ttccagacca
     2761 aagcttgctt caggaaaact gagtgggatc aatgaaatac ccaggccatt cagcccacct
     2821 gtaacttcca acaccagccc acctcctact gctcccttag cccgagccga gagcagctcc
     2881 tctatctcct cgtctgcctc gttgagtgct gccaacactc caacagtagg tgtatcacgg
     2941 ggtcccagcc cggtcagcct tggaaaccag gacactttac ctgtggcaat cgcccttaca
     3001 gaatctgtca acgcctactt taaaggagca gatcctacca agtgtattgt gaagatcacg
     3061 ggagacgtga caatatcatt tcccagtggg attattaaag tcttcaccag caacccctcc
     3121 ccagctgtgc tgtgctttag ggtgaagaac atcagcaggc tggagcagat ccttccaaac
     3181 tcccagcttg tgttcagtga tccatcacag tgtgattcca acacaaaaga tttttggatg
     3241 aacatgcaag ctgttactat ctacctcaag aagctatcag aacaaaatcc agctgcttct
     3301 tattataatg tagatgtatt aaaataccag gtatcatcaa atggcattca gtccactcct
     3361 ctaaatcttg caacctactg gaaatgtagt gccagcacca cagatctgag agtggattat
     3421 aagtacaacc cagaagctat ggtcgctccg agtgtgcttt ccaacataca ggtggtggtt
     3481 cccgtggatg gaggtgtcac aaatatgcag tcccttcctc ctgcaatatg gaatgcagag
     3541 caaatgaaag ctttttggaa attatctggg atttcggaga aatcagacag tggaggttct
     3601 ggatccctca gagcaaaatt tgatctttca gaaggaccca gcaaacctac aacactggca
     3661 gtgcaattcc tcagtgaggg caatacgctc tcaggagtgg atattgagct tgtgggtact
     3721 ggctatagac tttccttagt aaagaagcgc tttgccactg gacgatactt ggcagattgt
     3781 tgagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     3841 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3901 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3961 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4021 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     4081 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4141 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4201 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4261 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4321 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4381 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4441 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4501 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4561 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4621 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4681 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4741 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4801 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4861 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4921 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4981 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5041 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5101 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5161 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5221 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5281 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5341 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5401 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5461 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5521 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5581 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5641 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5701 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5761 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5821 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5881 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5941 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6001 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6061 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6121 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6181 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6241 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7141 atgcat
//
