LOCUS       sequence-410734-        7046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express tdMCP-msfGFP(V206K)-GBI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7046)
  AUTHORS   Hu Y, Xu J, Gao E, Fan X, Wei J, Ye B, Xu S, Ma W
  TITLE     Enhanced single RNA imaging reveals dynamic gene expression in live 
            animals.
  JOURNAL   Elife. 2023 Mar 3;12:e82178. doi: 10.7554/eLife.82178.
  PUBMED    36867026
REFERENCE   2  (bases 1 to 7046)
  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: "Elife."; 
            date: "2023-03-3"; volume: "12:e82178. doi"; pages: " 
            10.7554/eLife.82178"
COMMENT     Sequence Label: pcDNA-tdMCP-msfGFP(V206K)-GBI
FEATURES             Location/Qualifiers
     source          1..7046
                     /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"
     CDS             1715..2425
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1742..1770)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2365..2385
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2447..2608
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     primer_bind     complement(2637..2654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2643..2867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2913..3341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3000..3019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3210..3231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3350..3370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3355..3684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3535..3670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3597..3616
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3751..4545
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3805..3824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4415..4434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4719..4840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4756..4775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4810..4829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4889..4905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4889..4905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4902..4924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4913..4929
                     /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(4937..4943,4944..4961,4962..4967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4982..5003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5120..5137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5291..5879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5371..5390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6050..6841,6842..6910))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPKERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6673..6692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6911..7015)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aaggcttggt acctgctagc cgttaaaatg gcttctaact ttactcagtt
      961 cgttctcgtc gacaatggcg gaactggcga cgtgactgtc gccccaagca acttcgctaa
     1021 cgggatcgct gaatggatca gctctaactc gcgttcacag gcttacaaag taacctgtag
     1081 cgttcgtcag agctctgcgc agaatcgcaa atacaccatc aaagtcgagg tgcctaaagg
     1141 cgcctggcgt tcgtacttaa atatggaact aaccattcca attttcgcca cgaattccga
     1201 ctgcgagctt attgttaagg caatgcaagg tctcctaaaa gatggaaacc cgattccctc
     1261 agcaatcgca gcaaactccg gcatctacgc catggcttct aactttactc agttcgttct
     1321 cgtcgacaat ggcggaactg gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat
     1381 cgctgaatgg atcagctcta actcgcgttc acaggcttac aaagtaacct gtagcgttcg
     1441 tcagagctct gcgcagaatc gcaaatacac catcaaagtc gaggtgccta aaggcgcctg
     1501 gcgttcgtac ttaaatatgg aactaaccat tccaattttc gccacgaatt ccgactgcga
     1561 gcttattgtt aaggcaatgc aaggtctcct aaaagatgga aacccgattc cctcagcaat
     1621 cgcagcaaac tccggcatct acgcggattc tagaatggtg atgtacagtt caggaggcgg
     1681 cggaagcttg gatccaggtg gaggtggaag cggtagcaaa ggagaagaac ttttcactgg
     1741 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtccg
     1801 tggagagggt gaaggtgatg ctacaaacgg aaaactcacc cttaaattta tttgcactac
     1861 tggaaaacta cctgttccgt ggccaacact tgtcactact ctgacctatg gtgttcaatg
     1921 cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     1981 aggttatgta caggaacgca ctatatcttt caaagatgac gggacctaca agacgcgtgc
     2041 tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaagg gtattgattt
     2101 taaagaagat ggaaacattc ttggacacaa actcgagtac aactttaact cacacaatgt
     2161 atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa
     2221 cgttgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc caattggcga
     2281 tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctaagc tttcgaaaga
     2341 tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac
     2401 acatggcatg gatgagctct acaaaggtgg aggtcggacc gaagagtaca agcttatcct
     2461 gaacggtaaa accctgaaag gtgaaaccac caccgaagct gttgacgctg ctaccgcgga
     2521 aaaagttttc aaacagtacg ctaacgacaa cggtgttgac ggtgaatgga cctacgacga
     2581 cgctaccaaa accttcacgg taaccgaata agggcccgtt taaacccgct gatcagcctc
     2641 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2701 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2761 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2821 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2881 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3061 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3121 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3241 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3301 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3601 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3661 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3721 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3781 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3841 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3901 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3961 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4021 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4081 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4141 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4201 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4261 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4321 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4381 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4441 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4501 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4561 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4621 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4681 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4741 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4801 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4861 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4921 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4981 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5041 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5101 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5161 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5221 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5701 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6721 aacgttcttt ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7021 ttccccgaaa agtgccacct gacgtc
//
