LOCUS       sequence-347010-        4086 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 4086)
  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 4086)
  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: pDonorCm(Ptr)
FEATURES             Location/Qualifiers
     source          1..4086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     451..468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    585..606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(621..626,627..644,645..651)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    659..675
                     /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)."
     primer_bind     664..686
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     683..699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     683..699
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             831..1490
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(897..916)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(2169..2186)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2169..2185)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2178..2200)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2394..2413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2513..2535
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2573..2591)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2659..2763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2764..2832,2833..3624)
                     /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     complement(2982..3001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3795..4086,1..297)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
       61 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      601 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      661 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttttg
      721 ggtgttgttt gaagtataag ttgacatatc tgtactaaaa gatggcataa attggaagtg
      781 taaggtggca tagtctagta tttaaccaaa tggttaaatg gttgactcac atggagaaaa
      841 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
      901 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
      961 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1021 cccgcctgat gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga
     1081 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1141 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1201 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1261 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1321 acaacttctt cgcccccgtt ttcactatgg gcaaatatta tacgcaaggc gacaaggtgc
     1381 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1441 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     1501 gcagttattg gtgccctttt aattttcctt aattattttt aaagttagac tgatttagac
     1561 ttggaaaagc ttaatgattg gagagctaaa ttgactaata gtattcagtc aagtttaatt
     1621 agttttaagc gatccatgca taactatttc tgtacaatgc tatattttca ccaattaata
     1681 atttttaatc aagcctacat tatgaaatat actataccca tttgaactct tctatttgta
     1741 ccattgtcgg tagcaaaaac ttatggggtt ttgaacgtca cttaaattgt aagcatttgc
     1801 gatggaggcg cgtttagagt caaccttgat tctgatatgc tccgaatttt tggtaagaat
     1861 ataagtgtga gagtagctaa tgtggatacg cctgagttaa gggaaaaatg tgaaaatgaa
     1921 ataactcgtt atcatgcaaa gtgactaagg ttataatctt ccgtttatgg cacatagcag
     1981 ccaactaaac ttgacagtat ttttatgtgg ttggctttat aaaaccagca tttggtaaca
     2041 ttatgccaat ttttacttca atattatgcc aacatacact acactaacgg agctgtagca
     2101 caataagctc gtttgtactt atgccaactt atacttcaaa caacattggg ggtaccgagc
     2161 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2221 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2281 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     2341 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2401 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2461 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2521 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2581 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2641 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2701 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2761 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2821 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2881 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2941 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3001 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3061 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3121 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3181 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3241 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3301 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3361 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3421 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3481 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3541 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3601 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3661 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3721 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3781 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3841 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3901 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3961 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4021 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4081 tagtta
//
