LOCUS       sequence-409387-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human T6B peptide fused with mChery in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   Sekar V, Marmol-Sanchez E, Kalogeropoulos P, Stanicek L, Sagredo EA,
            Widmark A, Doukoumopoulos E, Bonath F, Biryukova I, Friedlander MR
  TITLE     Detection of transcriptome-wide microRNA-target interactions in 
            single cells with agoTRIBE.
  JOURNAL   Nat Biotechnol. 2023 Sep 21. doi: 10.1038/s41587-023-01951-0.
  PUBMED    37735263
REFERENCE   2  (bases 1 to 6338)
  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 
            Biotechnol. 2023 Sep 21. doi: 10.1038/s41587-023-01951-0."
COMMENT     Sequence Label: pcDNA3.1_T6B -mCherry
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..47)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      885..894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1158..1868
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1303..1321)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1571..1590
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1913..1930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1919..2143
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2189..2617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2276..2295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2486..2507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2626..2646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2631..2960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2811..2946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2873..2892
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3027..3821
                     /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(3081..3100)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3691..3710
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3995..4116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4086..4105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4165..4181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4165..4181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4178..4200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4189..4205
                     /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(4213..4219,4220..4237,4238..4243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4258..4279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4396..4413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4567..5155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4647..4666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5326..6117,6118..6186))
                     /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     5949..5968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6187..6291)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
       61 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      181 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      841 cgaaattaat acgactcact atagggagac ccaagctggc tagcgccacc atggattgtc
      901 aggctgtctt gcagactctt ttgagccgaa ctgatttgga ccccagggtg ctctcaaaca
      961 ctggctgggg ccaaactcaa attaagcagg acacagtgtg ggacattgaa gaggtgccaa
     1021 ggcctgaggg gaaatctgac aaaggaactg aggggtggga gagcgctgcc acacagacca
     1081 agaactcagg gggctgggga gatgcaccca gccaaagcaa tcaaatgaag tctggatggg
     1141 gggagctcgg aagcggaatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1201 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1261 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1321 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1381 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     1441 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1501 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1561 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1621 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1681 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1741 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     1801 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     1861 acaagtaggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     1921 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1981 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2041 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2101 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2161 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2221 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2281 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2341 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2401 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2461 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2521 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2581 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2641 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2761 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2821 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2881 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2941 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3001 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3061 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3121 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3181 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3241 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3301 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3361 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3421 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3481 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3541 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3601 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3661 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3721 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3781 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3841 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3901 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3961 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4141 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4201 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4261 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4321 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4981 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6301 ccgaaaagtg ccacctgacg tcgacggatc gggagatc
//
