LOCUS       sequence-351764-        3916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rox flanked gentamycin resistance cassette integrates into A2542 
            integration site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3916)
  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 3916)
  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: pCJ163
FEATURES             Location/Qualifiers
     source          1..3916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     591..610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     844..861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1390..1421
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    1422..1442
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             1557..2090
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRNSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     terminator      complement(2141..2168)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2184..2255)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2271..2302
                     /label=rox
                     /bound_moiety="Dre recombinase"
     promoter        2882..2986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2987..3055,3056..3847)
                     /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(3205..3224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
       61 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      241 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      721 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      781 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      841 cgcagcgagt cagtgagcga ggaagcggaa gaatgatcac gcggatatat tgggcgcgct
      901 tcggcacttc gatatcggcg agtttttcgg gggcattaac ggtgatcgct tcgttaaaca
      961 tccctgcggc gtagtcccaa cgactcacat aggggacata catgacattg gtgcgatttt
     1021 cggcaatttt ttccataccc cgatgcagat agccgatgac gggttcgcaa tcaacgacgt
     1081 cttccccgtc aagggtgacg atcaagcgta atacgccgtg catggagggg tggtgaggcc
     1141 ccatgttaat caccataggt tcggtacgag tttcaattct tgtcatggat ccgcaggtct
     1201 tccgttatgc ttatttaaat taaatctggt gatggggtga gatggtttgt ttgtgcattg
     1261 cgtttttcca atctcctgaa tcaccgttgc ttatgagttg cagtgattgg cttttagatg
     1321 gtttgacgct aggtttattt actctctgaa cattataaag tttcgcttaa ccatgacaag
     1381 cttgccccct aactttaaat aatgccaatt atttaaagtt actgcagcgg ccgctactag
     1441 tacaacaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1501 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatgt
     1561 tacgcaacag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     1621 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     1681 ctgctcttga tcttttcggt cgtgagttcg gagacgtagc cacctactcc caacatcagc
     1741 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     1801 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag tttgagcagc
     1861 cgcgtagtga gatctatatc tatgatctcg cagtctccgg cgagcaccgg aggcagggca
     1921 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     1981 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     2041 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa tcagaattgg
     2101 ttaattggtt gtaacactgg cagagcctct agtatataaa cgcagaaagg cccacccgaa
     2161 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
     2221 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggctctagt taactttaaa
     2281 taatgccaat tatttaaagt tattgggcga gaaaatggct gtgcttgatc cccgatgcag
     2341 gcttggtttg gtgtcgctgc gaaaaatttc gatacaatat agtgtgattt actttacaaa
     2401 accacacaat tcattgacgg cgaaaccact gtgacccagt atccttccgt tgcccccgac
     2461 cacaaacttg gttctaccac cttgacccgc tatgacgcca aggcgatcgc caaacattac
     2521 cgttggcgac cctggcaatc cctgtggcgg gcctttacga tcatttggtt atttgggtgg
     2581 ttttccttca aattgtggct agatagtaaa cgcgaagaag aacccgaaac ccgcgcccgt
     2641 cgcgccaacg aactgcgtga aattctgacc cgcttagggc cgacatttat caaagtaggg
     2701 caagcccttt caacccggcc agacttgatt cgtcccgact ttctcgaaga attaatcaag
     2761 ctccaagacc aactgccttc ttttgataac gagattgcct ttgatacgcc tatttttata
     2821 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2881 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2941 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3001 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3061 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3361 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3541 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3601 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3841 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3901 ttaatttaaa aggatc
//
