LOCUS       sequence-346302-        4606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA donor for transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4606)
  AUTHORS   Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S
  TITLE     Orthogonal CRISPR-associated transposases for parallel and 
            multiplexed chromosomal integration.
  JOURNAL   Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 
            10.1093/nar/gkab752.
  PUBMED    34478496
REFERENCE   2  (bases 1 to 4606)
  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. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752."
COMMENT     Sequence Label: pDonorGFP(Ptr)
FEATURES             Location/Qualifiers
     source          1..4606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(288..305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(422..967)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(502..521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1349..1371
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1379..2095
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1412..1440)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2035..2055
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2189..2237
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      2299..2347
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(2416..2438)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3290..3309)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3740..4399)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4314..4333
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4400..4502)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
       61 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
      121 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
      181 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
      241 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
      301 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
      361 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
      421 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
      481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
      541 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
      601 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
      661 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      721 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
      781 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
      841 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
      901 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
      961 atctcaagaa gatcatctta ttaatatacg tagtatctac gatacgtagt atctacgata
     1021 cgtaggaggt tgagaagtca tttaataagg ccactgttaa acgtagatat atctacgcgt
     1081 agatatatct acgatacgta gcggccgacg cgctgggcta cgtcttaatg ggcagcgccc
     1141 acatacgcag cgatttccaa cagcttgagg cggaagcttt aatgcgggtc ctttgggtgt
     1201 tgtttgaagt ataagttgac atatctgtac taaaagatgg cataaattgg aagtgtaagg
     1261 tggcatagtc tagtatttaa ccaaatggtt aaatggttga ctcaccggga attccatggt
     1321 cgacatgcat aagcttccta ggaataattt tgtttaactt taagaaggag atatacatat
     1381 ggctagcaaa ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg
     1441 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     1501 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     1561 tgtcactact ttcacttatg gtgttcagtg cttttcaaga tacccggatc atatgaaacg
     1621 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     1681 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     1741 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttgggcacaa
     1801 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     1861 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
     1921 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     1981 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     2041 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggatc
     2101 cggctgctaa tcgagttaat taaactagtg agctcggtac ccggggatcc tctagaggtc
     2161 gaaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct
     2221 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccaag ctaattcacc
     2281 tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct ttttttttgg
     2341 agattttcaa cgtgaaaaaa ttattattcg caattccaag ctctgcctcg cgcgtttcgg
     2401 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2461 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2521 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatgggctcg atcccctcgg
     2581 atccaattgg tcttaatttt ccttaattat ttttaaagtt agactgattt agacttggaa
     2641 aagcttaatg attggagagc taaattgact aatagtattc agtcaagttt aattagtttt
     2701 aagcgatcca tgcataacta tttctgtaca atgctatatt ttcaccaatt aataattttt
     2761 aatcaagcct acattatgaa atatactata cccatttgaa ctcttctatt tgtaccattg
     2821 tcggtagcaa aaacttatgg ggttttgaac gtcacttaaa ttgtaagcat ttgcgatgga
     2881 ggcgcgttta gagtcaacct tgattctgat atgctccgaa tttttggtaa gaatataagt
     2941 gtgagagtag ctaatgtgga tacgcctgag ttaagggaaa aatgtgaaaa tgaaataact
     3001 cgttatcatg caaagtgact aaggttataa tcttccgttt atggcacata gcagccaact
     3061 aaacttgaca gtatttttat gtggttggct ttataaaacc agcatttggt aacattatgc
     3121 caatttttac ttcaatatta tgccaacata cactacacta acggagctgt agcacaataa
     3181 gctcgtttgt acttatgcca acttatactt caaacaacat tggggtggat cctctacgcc
     3241 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     3301 gacatcaccg atggggaaga tcgggcctgc caccataccc acgccgaaac aagcgccctg
     3361 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
     3421 atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga ataaatgatc
     3481 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag
     3541 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     3601 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     3661 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     3721 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     3781 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     3841 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     3901 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     3961 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     4021 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     4081 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     4141 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     4201 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     4261 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     4321 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     4381 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     4441 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     4501 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     4561 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattc
//
