LOCUS       sequence-173231-        3893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gtbp‐1 (flanking sequence)::eGFP::recoded gtbp‐1(8/27bp, every 
            3bp)::PH::gtbp‐1 (flanking sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3893)
  AUTHORS   Paix A, Schmidt H, Seydoux G
  TITLE     Cas9-assisted recombineering in C. elegans: genome editing using in 
            vivo assembly of linear DNAs.
  JOURNAL   Nucleic Acids Res. 2016 Sep 6;44(15):e128. doi: 10.1093/nar/gkw502. 
            Epub 2016 Jun 1.
  PUBMED    27257074
REFERENCE   2  (bases 1 to 3893)
  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: "Nucleic 
            Acids Res."; date: "2016-09-6"; volume: "44(15):e128. doi"; pages: "
            10.1093/nar/gkw502. Epub 2016 Jun 1"
COMMENT     Sequence Label: AP1298‐1
FEATURES             Location/Qualifiers
     source          1..3893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(160..951,952..1020))
                     /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     783..802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1021..1125)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1193..1211
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1249..1271)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1371..1390
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1584..1606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1598..1615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1599..1615
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1672..1674,1675..1677,1678..1680,1681..2391)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MRVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     primer_bind     complement(1708..1736)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2331..2351
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2892..2908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2892..2908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2905..2927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2916..2932
                     /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(2940..2946,2947..2964,2965..2970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2985..3006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
       61 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      121 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      181 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      241 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      301 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      361 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
      421 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
      481 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      541 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      601 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
      661 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
      721 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
      781 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
      841 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
      901 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
      961 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1021 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1081 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1141 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1201 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1261 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1321 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1381 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1441 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1501 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     1561 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     1621 cgagctcggt acccggggat cggcggttcg ggtggtgctc cacgaggtgg tatgcgcgtg
     1681 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     1741 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
     1801 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
     1861 actactctga cctacggcgt gcagtgcttc agccggtacc ccgaccacat gaagcagcat
     1921 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
     1981 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     2041 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     2101 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     2161 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
     2221 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     2281 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
     2341 gagtttgtaa cagctgctgg gattacactc ggcatggacg agctgtacaa gagcggcttt
     2401 cagaacgccg gtcagaacca cgggctccag gatgacccgg accttcaggc ccttctgaag
     2461 ggcagccagc ttctgaaggt gaagtccagc tcgtggcgta gggaacgctt ctacaagcta
     2521 caggaggact gcaagaccat ctggcaggaa tctcgaaagg tcatgaggtc cccggagtcg
     2581 cagctgttct ccatcgagga cattcaggag gtacggatgg gacaccgcac agaaggcctg
     2641 gagaagtttg cccgagacat ccccgaggat cgatgcttct ccattgtctt caaggaccag
     2701 cgcaacaccc tagacctcat tgccccatca ccagctgacg ctcagcactg ggtgcagggc
     2761 ctgcgcaaga tcatccacca ctccggctcc atggaccagc ggcagaagta gaagctttcc
     2821 gttctccttt ttccttcttg taaggatcct ctagagtcga cctgcaggca tgcaagcttg
     2881 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2941 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3001 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc ttt
//
