LOCUS       sequence-488478-        2787 bp    DNA     circular SYN 06-OCT-2025
DEFINITION  Kanamycin/ColEI backbone with mScarlet-I placeholder for lvl2 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2787)
  AUTHORS   Inckemann R, Chotel T, Brinkmann CK, Burgis M, Andreas L, Baumann J,
            Sharma P, Klose M, Barrett J, Ries F, Paczia N, Glatter T, Willmund 
            F, Mackinder LCM, Erb TJ
  TITLE     Advancing chloroplast synthetic biology through high-throughput 
            plastome engineering of Chlamydomonas reinhardtii
  JOURNAL   bioRxiv 2024.05.08.593163
REFERENCE   2  (bases 1 to 2787)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.05.08.593163"
COMMENT     Sequence Label: pME_Cp_0_7-8_005_Kan/ColEI_mScarlet-I
FEATURES             Location/Qualifiers
     source          1..2787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             162..173
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      884..955
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      971..998
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1052..1867)
                     /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
                     ADRYQDLAILWKCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1775..1794
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1859..1878
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2092..2680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2581..2600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacatgagac gaggaaagtg aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc
       61 ttatttaata atgtgtgcgg caattcacat ttaatttatg aatgttttct taacatcgcg
      121 gcaactcaag aaacggcagg ttcggatctt agctactaga gaaagaggag aaatactaga
      181 tgcgtaaagg cgaagcagtg atcaaagaat ttatgcgctt caaagttcac atggaaggtt
      241 ctatgaacgg ccacgaattt gaaattgaag gtgaaggcga aggtcgtcca tacgaaggta
      301 ctcaaaccgc aaaactgaaa gttaccaaag gtggtccact gccattttct tgggatattc
      361 tgtctccaca atttatgtac ggttctcgtg catttatcaa acacccagca gatattccag
      421 actactacaa acaatctttt ccggaaggtt tcaaatggga acgtgttatg aattttgaag
      481 atggtggtgc agttacggtt acccaagata cctctctgga agatggtact ctgatctaca
      541 aagttaaact gcgtggtact aactttccac cagatggtcc agttatgcag aaaaaaacca
      601 tgggttggga agcatctacc gaacgtctgt acccagaaga tggcgttctg aaaggtgata
      661 tcaaaatggc actgcgtctg aaagatggcg gtcgttacct ggcagatttc aaaaccacct
      721 acaaagcgaa aaaaccagtt caaatgccag gtgcatacaa cgttgatcgt aaactggata
      781 ttaccagcca caacgaagat tacaccgttg ttgaacaata cgaacgttct gaaggccgtc
      841 actctaccgg tggtatggat gaactgtaca aataaccagg catcaaataa aacgaaaggc
      901 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
      961 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacgtctc aagactgcgg
     1021 ccgcgtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     1081 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1141 atgaaggaga aaactcaccg aggcatttcc ataggatggc aagatcctgg tatcggtctg
     1201 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1261 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     1321 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1381 catcaaccaa accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc
     1441 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1501 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     1561 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1621 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1681 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1741 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1801 aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat
     1861 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     1921 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     1981 gaataaatcg aacttttgct gagttgaagg atcagtcatg accaaaatcc cttaacgtga
     2041 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2101 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2161 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2221 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2281 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2341 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2401 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2461 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2521 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2581 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2641 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2701 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2761 tgattctgtg gataaccgtg cggccgc
//
