LOCUS       sequence-484593-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  24xGCN4::T2A with flanking restriction sites (KasI and AvrII). 
            Useful for cloning SunTag reporter plasmids for expression in C. 
            elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  TITLE     SunTag-based single-molecule translation imaging in C. elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4928)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSRG101 (KasI::24xGCN4::T2A::AvrII)
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     666..682
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     667..683
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             847..903
                     /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             919..975
                     /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             991..1047
                     /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             1063..1119
                     /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             1135..1191
                     /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             1207..1263
                     /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             1279..1335
                     /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             1351..1407
                     /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             1423..1479
                     /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             1495..1551
                     /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             1567..1623
                     /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             1639..1695
                     /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             1711..1767
                     /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             1783..1839
                     /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             1855..1911
                     /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             1927..1983
                     /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             1999..2055
                     /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             2071..2127
                     /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             2143..2199
                     /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             2215..2271
                     /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             2287..2343
                     /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             2359..2415
                     /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             2503..2559
                     /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             2614..2667
                     /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"
     primer_bind     complement(2687..2703)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2740..2758)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2779..2795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2779..2795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2792..2814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2803..2819
                     /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(2827..2833,2834..2851,2852..2857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2872..2893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3010..3027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3181..3769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3261..3280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3940..4731,4732..4800))
                     /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     4563..4582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4801..4905)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      661 ccccctcgag gtcgacggta tcgataagct tatgggcgcc atcatacacg cggccgcaat
      721 catcggtccg gatatcaagg tattttcctg catttttcaa ctgggaaaat gaaagaaaat
      781 cgataatttc aggaagatat caggcttatc ggtccgggtg gatctggagg tggaggttct
      841 ggaggagaag aacttttgag caagaattat catcttgaga acgaagtggc tcgtcttaag
      901 aaaggttctg gcagtggaga agaactgctt tcaaagaatt accacctgga aaatgaggta
      961 gctagactga aaaaggggag cggaagtggg gaggagttgc tgagcaaaaa ttatcatttg
     1021 gagaacgaag tagcacgact aaagaaaggg tccggatcgg gtgaggagtt actctcgaaa
     1081 aattatcatc tcgaaaacga agtggctcgg ctaaaaaagg gcagtggttc tggagaagag
     1141 ctattatcta aaaactacca cctcgaaaat gaggtggcac gcttaaaaaa gggaagtggc
     1201 agtggtgaag agctactatc caagaattat catcttgaga acgaggtagc gcgtttgaag
     1261 aagggttccg gctcaggaga ggaactgctc tcgaagaact atcatcttga aaatgaggtc
     1321 gctcgattaa aaaagggatc gggcagtggt gaggaactac tttcaaagaa ttaccacctc
     1381 gaaaacgaag tagctcgatt aaagaaaggt tcagggtcgg gtgaagaatt actgagtaaa
     1441 aattatcatc tggaaaatga ggtagcgaga ctaaaaaagg ggagtggttc tggcgaagag
     1501 ttgctatcga aaaattatca tcttgagaac gaagttgcta ggctcaaaaa gggctcaggc
     1561 tcaggcgagg agttgctctc gaaaaactac cacttggaaa atgaggtcgc gaggttgaaa
     1621 aaggggagcg ggtcgggcga ggagttattg agcaaaaact atcatttaga gaacgaagtc
     1681 gcgcgcttaa agaaaggctc gggctcgggc gaagaactct tatcgaagaa ctaccacctc
     1741 gaaaatgagg tcgccaggtt gaaaaagggc agtggcagcg gggaggaact cttgagcaag
     1801 aactaccact tggagaatga ggtcgcgaga ttgaagaaag ggtcggggag cggcgaggaa
     1861 ttgctcagca agaattatca tttggagaac gaagtcgcca ggctcaagaa aggctcgggg
     1921 tcgggggagg aattgttgag taaaaactac cacttggaaa atgaagtcgc caggctcaaa
     1981 aaagggagtg ggagcggcga agagttattg agcaaaaatt accacttgga gaacgaagtg
     2041 gcaaggctca agaaagggag cggcagcggg gaggagctct tatcgaagaa ctaccactta
     2101 gagaatgaag tcgcccgctt gaagaaaggc tcggggagcg gggaagagct cttgagcaag
     2161 aactaccact tggaaaatga ggtggcgcgc ttgaagaaag ggagcgggag cggggaagag
     2221 ttactatcta agaattatca tctcgagaac gaggtggctc gactaaagaa gggctccggc
     2281 agtggggagg aactcctgtc gaagaactat catcttgaaa atgaggttgc aagacttaaa
     2341 aaggggtccg gatcaggtga ggaactactc agtaagaatt accacctgga aaacgaagtt
     2401 gcacgtttga agaaaggatc aggatcaggc gaagaactgc tctcaaaaga ttatcatttg
     2461 gaaaatgagg ttgcacgttt aaaaaaggga agtggcagtg gtgaggaact tctgtcgaaa
     2521 aattatcatc tcgagaatga agtagcccga cttaaaaagg gttctggctc gggtcagcgg
     2581 ccgcaccggt ggtaccagga agctagctac gtagagggac gtggatccct cctcacctgc
     2641 ggagacgtcg aggagaaccc aggaccacct aggctgcagc ccgggggatc cactagttct
     2701 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaatttc
     2761 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3601 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3841 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3901 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3961 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4021 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4081 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4141 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4201 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4261 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4321 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4381 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4441 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4501 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4561 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4621 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4681 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4741 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4801 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4861 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4921 agtgccac
//
