LOCUS       sequence-430551-        6556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express tdMCP-6xGCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6556)
  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 6556)
  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-6xGCN4
FEATURES             Location/Qualifiers
     source          1..6556
                     /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             1687..1743
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1759..1815
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1831..1887
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1903..1959
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1975..2031
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             2047..2103
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(2147..2164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2153..2377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2423..2851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2510..2529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2720..2741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2860..2880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2865..3194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3045..3180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3107..3126
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3261..4055
                     /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(3315..3334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3925..3944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4229..4350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4266..4285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4320..4339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4399..4415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4399..4415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4412..4434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4423..4439
                     /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(4447..4453,4454..4471,4472..4477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4492..4513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4630..4647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4801..5389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4881..4900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5560..6351,6352..6420))
                     /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     6183..6202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6421..6525)
                     /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 aagcttggta cctgctagcc gttaaaatgg cttctaactt tactcagttc
      961 gttctcgtcg acaatggcgg aactggcgac gtgactgtcg ccccaagcaa cttcgctaac
     1021 gggatcgctg aatggatcag ctctaactcg cgttcacagg cttacaaagt aacctgtagc
     1081 gttcgtcaga gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt gcctaaaggc
     1141 gcctggcgtt cgtacttaaa tatggaacta accattccaa ttttcgccac gaattccgac
     1201 tgcgagctta ttgttaaggc aatgcaaggt ctcctaaaag atggaaaccc gattccctca
     1261 gcaatcgcag caaactccgg catctacgcc atggcttcta actttactca gttcgttctc
     1321 gtcgacaatg gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc
     1381 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt
     1441 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg
     1501 cgttcgtact taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag
     1561 cttattgtta aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc
     1621 gcagcaaact ccggcatcta cgcggattct agaatggtga tgtacaaggg tggaggttct
     1681 ggaggagaag aacttttgag caagaattat catcttgaga acgaagtggc tcgtcttaag
     1741 aaaggttctg gcagtggaga agaactgctt tcaaagaatt accacctgga aaatgaggta
     1801 gctagactga aaaaggggag cggaagtggg gaggagttgc tgagcaaaaa ttatcatttg
     1861 gagaacgaag tagcacgact aaagaaaggg tccggatcgg gtgaggagtt actctcgaaa
     1921 aattatcatc tcgaaaacga agtggctcgg ctaaaaaagg gcagtggttc tggagaagag
     1981 ctattatcta aaaactacca cctcgaaaat gaggtggcac gcttaaaaaa gggaagtggc
     2041 agtggtgaag agctactatc caagaattat catcttgaga acgaggtagc gcgtttgaag
     2101 aagggttccg gctcaggata agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2341 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2401 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2641 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2701 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2761 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2821 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2881 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2941 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3001 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3061 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3121 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3181 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3241 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3301 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3361 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3421 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3481 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3541 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3601 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3661 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3721 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3781 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3841 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3901 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3961 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4021 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4081 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4141 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4201 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4321 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4381 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6541 agtgccacct gacgtc
//
