LOCUS       sequence-394740-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pACYC-plac-MycaTnsABC, lac promoter expressing transposase genes 
            TnsABC of McCAST (Tn7575)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6110)
  AUTHORS   Hsieh SC, Peters JE
  TITLE     Discovery and characterization of novel type I-D CRISPR-guided 
            transposons identified among diverse Tn7-like elements in 
            cyanobacteria.
  JOURNAL   Nucleic Acids Res. 2023 Jan 25;51(2):765-782. doi: 
            10.1093/nar/gkac1216.
  PUBMED    36537206
REFERENCE   2  (bases 1 to 6110)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2023-01-25- 25"; volume: "51"; issue: 
            "2"; pages: "765-782"
COMMENT     Sequence Label: pOPO635
FEATURES             Location/Qualifiers
     source          1..6110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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)."
     RBS             248..259
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4173..4199
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4256..4272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4256..4272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(4452..4997)
                     /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(4532..4551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5105..5199)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5220..5879)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5794..5813
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5880..5982)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     6069..6091
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6083..6100
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6084..6100
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccgcacgcat ctggaataag gaagtgccat tccgcctgac ctgcgcccaa tacgcaaacc
       61 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcatgaaaaa gaggagaaag gatctatgct tgacgcaaag agttttgata actggtgtca
      301 gagtcttggt gttagtcaaa caacccgtgc cgtcattgaa ggggttcgca actctgctcc
      361 ttcacgtcga gtgcagagcc gtcggggaaa tgttgttggc aattacccta gccacaaaat
      421 gggctttacc attcaatttg aatcacaccg caacgagtta gcgcgaatat atgagctaga
      481 gcatgattcc aacgttttgg agtattacga ccaacctata cctattgagc ttgactacct
      541 atccaagaat gggcggcgta accggcatcc atacacacca gattttttca tcatccgaaa
      601 agattctgcg ggttgggagg aatgcaagac tgagcaagag cttatcaagt taatagagga
      661 tagccccaac cgttatcaac aagatgagca gagccaatgg cattgtcctc ccgctgaaac
      721 ctacgcagca cgatatggct tgttttttaa agtacggtca gattccgaaa taaactggat
      781 ttttcaacaa aacttcgctt ggctggaaga ttattttcac ctagatagtt cagagcttag
      841 tgaagaagct gtgcaaaacc tgcttaagca ggtttccgat gaagagggca ttacgctagc
      901 agaattactc cggcaagcta acccagacga tatctatacc ttaattgcaa ctgactcact
      961 gtatgtcgat ctccatgctg ctcgattagc agaacccgat cgcgtccaag tcttttctga
     1021 tcaagcaact gcctttgctt atcagcgtgc tacagaagtt gctctatccc ttccaaatca
     1081 ctcacaaatc atttctataa cagttggggc ttctcttgtt tgggatggcg agtgctggga
     1141 agtcgttaat actggaagta ctacaaccgg actacttaga gcagataacc agttcgtaga
     1201 gctaccaaat ggaaccgttg aaagtctcat tcagcaaggc aaaattactg ccctaaccga
     1261 aactgaagac acaactcgtc aggacaaaat tcaagaaatt ctgcggcgtg ctaagcctgg
     1321 ggacattgcc gaagcaaatc gaaggtatga gcaaattcag ccttatctca acccagatgc
     1381 tccacctaca ccttctagca ccattcgtcg ttggcgaatt cagtaccgtc aagcagaggc
     1441 agcctatggc agtggatatg tcggactcct accaattaat tatgcaaaag ggaaccgaac
     1501 gagcaaactt ggtgagcctg ttcgagaatt catgattaaa tttatcaagg aaaactacga
     1561 aaccctgaag catcgtagaa ggtggcgagt ctacgagtct ttagttgctg cttgtgaagc
     1621 ccatcaacct aaacttactc ctcctagttc tacaaccttc tataaggaga tcaagcgccg
     1681 ctccggcatg gaacaaacgg caaaacggga agggcgacgg gcagctatcc aggaacgtcc
     1741 tacttatttg cagttggaat taaccactcc tcgacatggc gatcgcccct ttgaaattat
     1801 tcacattgac cataccgaaa ttgacgttga actggctatt tcgttgactt cccttactca
     1861 atgcaatttc aactccggta attcaggtct tcctaattta ggaagacctt gggcgacatt
     1921 catggttgat gcttatagcc gtcgtctctt agcagtctat ttgagttacg aggaacccag
     1981 ctaccgttcc tgcatgtccg tattgcgtat ctgtgttcaa cgtttcaaac gatttcctca
     2041 gtcaatcgtg gtagataatg gcccagaatt ccacagcaac tattttgaac aactactagc
     2101 atattttgct tgcacgaaaa aacaccgtcc tccagcagat gctagatatg gctttgtcgt
     2161 agagcgtttg tttggtactg ctaacactca atttgttcat gagttacgtg gtaatactca
     2221 aattaaacgt ttgcaccgca aagttacaaa atctgtcaaa cctgagagcc aggcagtttg
     2281 gacactgggt gagttgtatg aagctttctg ccactgggca tatgaagtct acgataagcg
     2341 acagcatcca acactaggac aaagcccccg tgaagccttc aatgatggtt tagccattgg
     2401 cggtagccga acttgccgcc gagtggaata tgacaaaacc ttccgaattc tgactttgcc
     2461 agctccagaa cgaggaaacc ggaaggtaca accggggcga ggcatcaaaa ttaataacat
     2521 ttactattgg tcgaatactt tccgtgaccc agaaattgag cagtcaaagg ttgaagtgcg
     2581 gtatgaacct tttgatgcca gtattgccta cgcattggtg cgggggcagt gggtggagtg
     2641 tgtttctgat tgctaccagt atcttcaggg acgttccgaa aaagaaatcc gtttggtcag
     2701 tgcagaatta gtaaaacgta agcgagatca aggacggaat gtgatcacct ccgacaagga
     2761 actggtgcag tttttaaact ctgttgaagc caaggaaggt aagtttttgg aacaacgcct
     2821 aaaagcttct gagaacaagc aagtagtgca ggcaattaca gctgttgaaa ctgattgctc
     2881 tgttgattca gtagttcctt cgagtcctgt tactgcagat ggtctatcag atagggaacc
     2941 gctgcttgat gaacgactag agacaaatga gcctaattct gatagtagtt ctagcaattt
     3001 ggaattctac ggagagtggt aattatgcag ttccaaaaag gctttccgcg agagttatta
     3061 aatcaaccca ttgctgatag agttgcctat tttcagagct atacaatgcc tcatcctaag
     3121 cttaaagatg ccgctcaaca gttgatgctc tcaattcatc aacctgctgg agcctctcta
     3181 atctttgtct ttggaccaac aggggttggc aaatccacct tattaaggaa ggttagaaaa
     3241 gaactaattg aagcagcgtt acctcggatg gaaattgata aggggtatat tccagttgcc
     3301 ggaatcgaag cgatcgctcc agaatttagc aattttgatt ggaaagattt ttatgtacgc
     3361 ggacttatgg ctcttaatga gccgatgatt gacaagaaga ttaatcgtac caattcaaag
     3421 cttaaacttc gtttggcatt agagtctgcc ctaagcaatc gaaagcctga tgcatttttt
     3481 gtcgatgagg cgcaaaattt gggcaaggtg gtgagcggac gcaagctacg tgatcaaact
     3541 gactgcatca aatccctcgc aaatattgct caaacccggt ttgttctttg tggcacatat
     3601 gagcttttaa tgctacggaa cttgagtgct caattgtgca gacgtagtac tgatattcat
     3661 ctccctcgct actatgcaga atctactcag gatcgggaaa cgtttcaggg cattgtgcaa
     3721 accctccagt tttaccttcc cctgccagaa actcccgatt tactgggttt atgggagttt
     3781 tgttatgaac gtagtattgg ctgtgtgggc attctgaagg actggctatc acgcacctta
     3841 aatgctgttc tggaaaaaga tccaaatgca gctactttaa cagctgaaga tttggaacat
     3901 catgcttggt caatggagca atgcatgatt atgctatccg aagcaaagga agaggagcag
     3961 aagttgacgc ctagagcaga aattcggaac gaactgcgtg ccagcctagg attagattta
     4021 cttgcttcgt ctaaagcgga aagtcccacc gaacaaaaaa aatccagaac aaagaaaacc
     4081 caagacatag gcaaacctct cccccaacgc cgccctgttg ggaaaaacca atatgctaat
     4141 ccatgataac aatgccaggc atcaaattaa gcagaaggcc atcctgacgg atggccttta
     4201 ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat
     4261 agctgtttcc tgtgtgaaat tgttatcctc tcagagggca caatcctatt ccgcgctatc
     4321 cgacaatctc caagacatta ggtggagttc agttcggcga gcggaaatgg cttacgaacg
     4381 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     4441 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     4501 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     4561 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     4621 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     4681 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     4741 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     4801 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     4861 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     4921 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     4981 gaccaaaacg atctcaagaa gatcatctta ttaagtctga cgctctattc aacaaagccg
     5041 ccgtccatgg gtagggggct tcaaatcgtc cccccatacg atataagttg ttactagtgc
     5101 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     5161 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
     5221 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     5281 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     5341 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     5401 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     5461 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     5521 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     5581 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     5641 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
     5701 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     5761 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     5821 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     5881 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     5941 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttgatacctg
     6001 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     6061 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat
//
