LOCUS       sequence-374490-        5463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rbcLXS7002 integrative plasmid expresses DRE recombinase as a 
            translational fusion to rbcL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5463)
  AUTHORS   Jones CM, Parrish S, Nielsen DR
  TITLE     Exploiting Polyploidy for Markerless and Plasmid-Free Genome 
            Engineering in Cyanobacteria.
  JOURNAL   ACS Synth Biol. 2021 Sep 17;10(9):2371-2382. doi: 
            10.1021/acssynbio.1c00269. Epub 2021 Aug 17.
  PUBMED    34530614
REFERENCE   2  (bases 1 to 5463)
  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: 
            "10.1021/acssynbio.1c00269"; journalName: "ACS Synth Biol"; date: 
            "2021-09-17- 17"; volume: "10"; issue: "9"; pages: "2371-2382"
COMMENT     Sequence Label: pCJ164
FEATURES             Location/Qualifiers
     source          1..5463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      119..707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     608..627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     861..878
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1584..2606
                     /product="site-specific recombinase"
                     /label=Dre recombinase
                     /note="Dre is a site-specific recombinase from 
                     bacteriophage D6. Recombination occurs at rox sequences."
     primer_bind     complement(2679..2697)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2693..2740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2875..2894)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2886..3701
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2886..3701
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2959..2978)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(3752..3779)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3795..3866)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        4446..4550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4551..4619,4620..5411)
                     /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(4769..4788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
       61 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      241 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      361 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      421 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      481 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      541 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      601 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      661 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      721 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      781 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      841 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag caggatcccc
      901 tccaactgaa acaccaaaca ccgaagtgat gccgacggtt ccagaatcaa aggaatccct
      961 agaatcgggc gaaaaaacac cagttgtcgg gcaagtttat atcaatcaac tattaatgac
     1021 cctattccct caccaaaatt ctctcaatac cccaaatcaa ccagatgagc cttagcccca
     1081 caaaactttc atgattgctt gagtgaaaat taaatgttta aagttcttaa aggagattgt
     1141 tgagaaacat aaatacttaa ttcatctcat ttgaacgctt tccctctcta aagatcccga
     1201 cagaaaacgg ttttagccca atgtctcatt aggtagcatg gctgaattcg agcgggattt
     1261 tatggctttt ttaggtattt ttgtaagggt aaaataggcc catcaaacag cattagaaat
     1321 gctaatcagc ccaaaaaaca aaagcaatct ttttttgttg ctaaaagata aaaataagtc
     1381 gaggctgtgg taacatatcc cacagattaa agaaagtcat aagacttgaa tcttcagaat
     1441 tttaaaaagc agttttgcca acgtaagatt tttgaagttt tcgaccaaca ataccgttac
     1501 tggtatttgt ctgttaaaga taagcatttt tgctggagga aaaccgcatg gttcagacca
     1561 aatctgctgg gtttaatgcc ggttctgagc tgattattag tggttcatct ggtggattcc
     1621 tgcgaaacat cggcaaagag tatcaggagg ccgctgaaaa cttcatgagg tttatgaatg
     1681 accagggggc gtacgctcct aacactttga gggatttgag gttggtcttt catagctggg
     1741 ccagatggtg ccatgctcgg cagcttgcat ggtttccaat tagtcctgaa atggcacgcg
     1801 aatactttct tcagttgcac gatgcagacc tggcctccac taccatcgac aagcactatg
     1861 ctatgcttaa tatgcttctg tcccactgcg gactgccacc cttgtccgac gacaagtcag
     1921 tgagtcttgc catgagaaga attagaagag aagccgcaac cgaaaagggt gagaggacag
     1981 gacaggcaat ccccctgcgc tgggacgacc tgaagctgct ggatgtgctg ctcagcagga
     2041 gcgagcggct ggtcgacctg cgcaacaggg ctttcctgtt cgtagcctat aacaccctca
     2101 tgagaatgtc tgaaatatca cgcatcaggg ttggggactt ggatcagaca ggagacacag
     2161 tgaccctgca catcagtcac actaagacaa tcaccacagc tgcgggcctt gacaaagtgc
     2221 tctcccggcg aaccacagca gtgctcaatg actggctgga cgtcagtggg cttagagaac
     2281 atccagacgc tgtgctcttc ccacctatac accggtcaaa caaagcccgc attactacca
     2341 cgcccctgac cgcccctgcc atggagaaga ttttcagtga tgcctgggtg ctgctgaaca
     2401 aacgggacgc cacccccaat aaagggaggt ataggacctg gaccggccat tccgccaggg
     2461 tgggtgccgc aatagacatg gccgagaaac aggtgtctat ggtcgagatt atgcaggaag
     2521 ggacatggaa gaagcctgaa acactgatgc ggtatctcag aaggggcgga gtgtccgtgg
     2581 gagccaattc tcgactgatg gatagctgac gagggcggtg ctttggcagg atccggctgc
     2641 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     2701 accccttggg gcctctaaac gggtcttgac gggttttttg tctagatcaa cggcctcaac
     2761 ctactactgg gcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     2821 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2881 gtgttatgag ccatattcaa cgggaaacgt cttgctccag gccgcgatta aattccaaca
     2941 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     3001 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3061 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3121 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3181 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     3241 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     3301 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3361 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3421 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     3481 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3541 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3601 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3661 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3721 tgtaacactg gcagagcctc tagtatataa acgcagaaag gcccacccga aggtgagcca
     3781 gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3841 ttcgactgag cctttcgttt tatttgggtt ttgttggttt ttgtgacctg aatttaattt
     3901 gaatcatgcg gaatgcgatc gccttaggac ggtcgcattt tttgtttacg tctaaaatta
     3961 gtcgaaatcc cccatcaacg ccatggtagt gattttgtct taacgttatt cacccatcaa
     4021 tttcaaaatg aacctgaagt ttcttaaatc tctctgggct acggcggcga tcgcctttgc
     4081 cattagcgta aatccgagcc ttgtctttgc tgaaacggaa cccccaagcg aaaccaaaac
     4141 tgccctgatc aatgaactcc gcaccttaac tttcctggat gaaaacgcga cccagatttt
     4201 ggacttgatg ctgcaacaga tccaagctca atccaccacc atgggcgatg gctttttggg
     4261 tgaagaaaca gatcccgaaa ccctggccgc tatccaagaa agtgtcaccc gcatcaccga
     4321 ccgcatttat accctgatgc aagacagaat tgatttcgtt gccctcgata cgcctatttt
     4381 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4741 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4921 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5401 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5461 att
//
