LOCUS       sequence-173227-        4560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gtbp‐1 (~500 bp flanking sequence)::GFP (with 3 introns)::gtbp‐1 
            (~500 bp flanking sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4560)
  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 4560)
  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: AP553‐1
FEATURES             Location/Qualifiers
     source          1..4560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(166..754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(246..265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(925..1716,1717..1785))
                     /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     1548..1567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1786..1890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1958..1976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2014..2036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2136..2155
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2349..2371
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2363..2380
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2364..2380
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2938..2966)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3714..3734
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4324..4340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4324..4340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4337..4359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4348..4364
                     /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(4372..4378,4379..4396,4397..4402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4417..4438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4555..4560,1..12))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
       61 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1981 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2041 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2101 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2161 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2221 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     2281 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     2341 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacccg
     2401 gggatcgcgc caagctcagg aaagttcaga tgccaaccta aatgagcgta agatctattt
     2461 gggaggaatc actcgagaga ttgtccccag caacacagct caggctgaat cggaaatcaa
     2521 gcaggtgttc tacagtaagc aaattaatag tcccaatagt tcactaaatt ttcattttaa
     2581 gaatatggag aaatcgagga ggttcgcatg ccgcgcaagg ttcttgacag tccgaacgac
     2641 acatcgaaac atgcctttgc atttattacc atgcgtaaca aggccggagt aaaagccatt
     2701 ttcgaagcgg ctacaaagga tgagcaaggc gtgtggaagc ttccactcaa actggatgca
     2761 ttcggatttg acggacatgc tcaaattagt gagcaacgtg atcaaccaca aggcggcgga
     2821 ttccgaggag gacgtggagg tttccaacca cgcggtggac aattcttccg ccgtggcggt
     2881 tcgggtggtg ctccacgagg tggtatgcgc agtaaaggag aagaactttt cactggagtt
     2941 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     3001 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
     3061 aaactacctg ttccatgggt aagtttaaac atatatatac taactaaccc tgattattta
     3121 aattttcagc caacacttgt cactactttc tgttatggtg ttcaatgctt ctcgagatac
     3181 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     3241 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtaagtt taaacagttc
     3301 ggtactaact aaccatacat atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat
     3361 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     3421 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     3481 aagaatggaa tcaaagttgt aagtttaaac atgattttac taactaacta atctgattta
     3541 aattttcaga acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     3601 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     3661 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     3721 cttgagtttg taacagctgc tgggattaca catggcatgg acgaactata caaaagcggt
     3781 ttccaaaatg cgggacaaaa ttagaagctt tccgttctcc tttttccttc ttgtaagaat
     3841 tgcacatcca ttagattctg tacctatcct tctcattttc ctatttattg tttctccttt
     3901 ccttcctcac cgttctaata atgatgattt aagtaagaag attctggtat ctgaatttta
     3961 tggcgtgtag cgttttagta gatctctcaa caatccaatc caatcagccc gcatcccatt
     4021 ttcacatcct atctccccgg ttttcggtga ttgacgaccg tgatcccatc tgagtttact
     4081 cttttcgagt acaagttgca atcagacatc atcccaccac cgaatatcca agttgtattt
     4141 gaatgttgtt ttaatatatt accactttcc ttctcctttt acaagccccc gttattgttt
     4201 ccttcattgg tgttcgggag tactctttgt actcaagttc ttacccccat tttggcacgt
     4261 tggaattaaa ataaaggatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc
     4321 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4381 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4441 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4501 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
//
