LOCUS       sequence-335053-        3116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Viral T2A linker element for assembly of TL3 to TL5 parts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3116)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3116)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pT2A
FEATURES             Location/Qualifiers
     source          1..3116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..37)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(191..779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(271..290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(950..1741,1742..1810))
                     /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     1573..1592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1811..1915)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1993..2448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2130..2151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2342..2361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2460..2909)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDWAMSG
                     LASRVRGSPPHLRRSAGFRGPPTRPLIAQSNSRGRHGGREHATSGPIRPIVSRITIHWP
                     SFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     2574..2596
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2588..2605
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2589..2605
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2612..2631
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2612..2630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2661..2670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2665..2674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2711..2764
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     promoter        complement(2869..2887)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2870..2887)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2905..2921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2905..2921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2918..2940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2929..2945
                     /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(2953..2959,2960..2977,2978..2983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
       61 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      121 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      841 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      901 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1021 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1081 tgataccgcg cgacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1141 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1201 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1261 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1321 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1381 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1441 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1501 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1561 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1621 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1681 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1741 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1801 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1921 ttccccgaaa agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     1981 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2041 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2101 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2161 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgctggc ccactacgtg
     2221 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2281 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2341 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2401 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     2461 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2521 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2581 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg
     2641 cccgacgtcg catgctcccg gccgccatgg cggccgcggg aattcgattg ggcgatgagt
     2701 ggtctcgtcg gagggccgcg gaagcctgct gacctgcgga gatgtggagg agaaccccgg
     2761 acccggctcg cgagaccact catcgcccaa tcactagtga attcgcggcc gcctgcaggt
     2821 cgaccatatg ggagagctcc caacgcgttg gatgcatagc ttgagtattc tatagtgtca
     2881 cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3061 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgta
//
