LOCUS       sequence-351757-        5466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rbcLXS7002 integrative plasmid expresses CRE recombinase as a 
            translational fusion to rbcL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5466)
  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 5466)
  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: pCJ111
FEATURES             Location/Qualifiers
     source          1..5466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        516..620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(621..689,690..1481)
                     /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(839..858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1652..2240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2141..2160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2394..2411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4215..4233)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4229..4276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4411..4430)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4422..5237
                     /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             4422..5237
                     /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(4495..4514)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(5288..5315)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5331..5402)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggtcgcattt tttgtttacg tctaaaatta gtcgaaatcc cccatcaacg ccatggtagt
       61 gattttgtct taacgttatt cacccatcaa tttcaaaatg aacctgaagt ttcttaaatc
      121 tctctgggct acggcggcga tcgcctttgc cattagcgta aatccgagcc ttgtctttgc
      181 tgaaacggaa cccccaagcg aaaccaaaac tgccctgatc aatgaactcc gcaccttaac
      241 tttccgggat gaaaacgcga cccagatttt ggacttgatg ctgcaacaga tccaagctca
      301 atccaccacc atgggcgatg gctttttggg tgaagaaaca gatcccgaaa ccctggccgc
      361 tatccaagaa agtgtcaccc gcatcaccga ccgcatttat accctgatgc aagacagaat
      421 tgatttcgtt gccctcgata cgcctatttt tataggttaa tgtcatgata ataatggttt
      481 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      541 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      601 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      661 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      721 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      781 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      841 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      901 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      961 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1021 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1081 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1141 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1201 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1261 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1321 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1381 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1441 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1561 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2281 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2341 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2401 cagtgagcga ggaagcggaa gagcaggatc ccctccaact gaaacaccaa acaccgaagt
     2461 gatgccgacg gttccagaat caaaggaatc cctagaatcg ggcgaaaaaa caccagttgt
     2521 cgggcaagtt tatatcaatc aactattaat gaccctattc cctcaccaaa attctctcaa
     2581 taccccaaat caaccagatg agccttagcc ccacaaaact ttcatgattg cttgagtgaa
     2641 aattaaatgt ttaaagttct taaaggagat tgttgagaaa cataaatact taattcatct
     2701 catttgaacg ctttccctct ctaaagatcc cgacagaaaa cggttttagc ccaatgtctc
     2761 attaggtagc atggctgaat tcgagcggga ttttatggct tttttaggta tttttgtaag
     2821 ggtaaaatag gcccatcaaa cagcattaga aatgctaatc agcccaaaaa acaaaagcaa
     2881 tctttttttg ttgctaaaag ataaaaataa gtcgaggctg tggtaacata tcccacagat
     2941 taaagaaagt cataagactt gaatcttcag aattttaaaa agcagttttg ccaacgtaag
     3001 atttttgaag ttttcgacca acaataccgt tactggtatt tgtctgttaa agataagcat
     3061 ttttgctgga ggaaaaccgc atggttcaga ccaaatctgc tgggtttaat gccggttcca
     3121 acctgctgac ggtgcaccag aatctgccgg cactgccggt cgatgcaacc agtgacgaag
     3181 tgcgcaaaaa tctgatggat atgtttcgtg accgccaagc cttcagcgaa catacgtgga
     3241 aaatgctgct gtcggtttgc cgtagctggg cggcctggtg taaactgaac aatcgcaaat
     3301 ggtttccggc agaaccggaa gatgtgcgtg actatctgct gtacctgcag gcacgtggtc
     3361 tggcagttaa aaccatccag caacatctgg gccaactgaa catgctgcac cgtcgctctg
     3421 gtctgccgcg tccgagtgat tccaatgccg tcagtctggt gatgcgtcgc attcgtaaag
     3481 aaaacgtgga tgcaggcgaa cgcgctaaac aggcactggc ttttgaacgt accgatttcg
     3541 accaagttcg ctctctgatg gaaaacagtg atcgttgcca ggacatccgc aatctggcat
     3601 tcctgggtat tgcttataac accctgctgc gcattgcaga aatcgctcgt attcgcgtga
     3661 aagatatcag ccgtacggac ggcggtcgca tgctgattca catcggccgt accaaaacgc
     3721 tggtttccac cgcgggcgtc gaaaaagccc tgtcactggg tgtcacgaaa ctggtggaac
     3781 gctggatttc agtttcgggc gtcgcagatg acccgaacaa ttacctgttt tgtcgtgtgc
     3841 gcaaaaatgg tgttgcagct ccgagcgcta cctctcagct gagtacgcgt gcgctggaag
     3901 gcatcttcga agccacccat cgcctgattt atggcgcgaa agatgacagc ggtcagcgtt
     3961 acctggcatg gtccggtcac tcagctcgtg ttggtgcagc acgtgatatg gcacgtgcag
     4021 gtgtctctat cccggaaatt atgcaggccg gcggttggac gaacgtgaat attgttatga
     4081 actatattcg taacctggac tctgaaacgg gtgcgatggt gcgtctgctg gaagatggcg
     4141 actgacgagg gcggtgcttt ggcaggatcc ggctgctaac aaagcccgaa aggaagctga
     4201 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     4261 cttgacgggt tttttgtcta gatcaacggc ctcaacctac tactgggcaa caaagccacg
     4321 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     4381 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     4441 aaacgtcttg ctccaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     4501 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     4561 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     4621 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     4681 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat
     4741 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     4801 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     4861 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     4921 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc
     4981 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     5041 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     5101 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     5161 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     5221 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcctctagt
     5281 atataaacgc agaaaggccc acccgaaggt gagccagtgt gactctagta gagagcgttc
     5341 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     5401 tgggttttgt tggtttttgt gacctgaatt taatttgaat catgcggaat gcgatcgcct
     5461 taggac
//
