LOCUS       sequence-479137-        4299 bp    DNA     circular SYN 01-AUG-2025
DEFINITION  Expresses TCTN3 minigene 
            (EFS-6xMyc-TCTN3-Ex10-I10-Ex11-copGFP-3xFLAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4299)
  AUTHORS   Che R, Panah M, Mirani B, Knowles K, Ostapovich A, Majumdar D, Chen 
            X, DeSimone J, White W, Noonan M, Luo H, Alexandrov A
  TITLE     Identification of Human Pathways Acting on Nuclear Non-Coding RNAs 
            Using the Mirror Forward Genetic Approach.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 19:2024.09.26.615073. doi: 
            10.1101/2024.09.26.615073.
  PUBMED    39386709
REFERENCE   2  (bases 1 to 4299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Jan 19:2024.09.26.615073. doi: 
            10.1101/2024.09.26.615073."
COMMENT     Sequence Label: TCTN3 minigene (pAVA3939)
FEATURES             Location/Qualifiers
     source          1..4299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..19)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2..19)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(39..56)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    45..269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    310..331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(448..465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(619..1207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(699..718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1378..2169,2170..2238))
                     /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     2001..2020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2239..2343)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2447..2658
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2727..2756
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2766..2795
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2805..2834
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2844..2873
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2883..2912
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2946..2975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3521..4186
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             3521..4186
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             4214..4285
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
ORIGIN
        1 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
       61 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
      121 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
      181 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
      241 gcatgctggg gatgcggtgg gctctatggc tctgacctat cgcggtcgtc ttaatgcctg
      301 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      361 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2341 gcgcacattt ccccgaaaag tgccacctga cgtcattaag gatattcacc attatcgtca
     2401 tttaaatagt cgcaaacgcg actgttcggc tccggtgccc gtcagtgggc agagcgcaca
     2461 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2521 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2581 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2641 tttgccgcca gaacacaggt gtcgtgacgc gaatacgact cactataggg agacccaagc
     2701 ttggatccca tcgatttaaa gctatggagc aaaagctcat ttctgaagag gacttgaatg
     2761 aaatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa aagctcattt
     2821 ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac ttgaatgaaa
     2881 tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagagcttg ggcgacctca
     2941 ccatggagca aaagctcatt tctgaagagg acttgaattt aagatctatg gcttttcaac
     3001 agagcacagc tgcttctctc accagtccta gaagtgggaa tcctggctat atagttggga
     3061 agccactctt ggctctgact gatgatataa gttactcagt atccttttta gagctgggtg
     3121 gcctgttgca gcctaatgag aaaagctgca aaggctttca aacttatgtt agactagcca
     3181 aaggtgagga attttttgtt cattataatg aggtacttat atactaatat atggttaatc
     3241 atttttggaa tctagttgtc tctctgtggt tttctgagga aaaaaatcac aaatttgtga
     3301 ctcgaacatt atgatagtaa tacaaaataa atagcattaa aggagaatga gaacataaat
     3361 tttgcaatgc tgcctccttt ttttttctgt tatgatgtcc ttgatcatca cttttccacc
     3421 ttggatgacc ctcttacaga gccagggtaa tggaagttgc tctgttaaaa gacatgaagt
     3481 gcagtttgga gtgaatgcaa tatctggatg caagctcagg atgcccgcca tgaagatcga
     3541 gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg
     3601 cacccccgag cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt
     3661 cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc
     3721 cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg
     3781 catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc
     3841 cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat
     3901 cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga
     3961 taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag
     4021 cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa
     4081 cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg
     4141 catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat ctcgagctcg
     4201 aatcgatgtc gaggattaca aggatgacga cgataaggac tataaggacg atgatgacaa
     4261 ggactacaaa gatgatgacg ataaataggg cccggtacc
//
