LOCUS       sequence-476154-        2883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ampicillin/ColEI backbone with mScarlet-I placeholder for lvl1 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2883)
  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 2883)
  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"; journalName: "bioRxiv 
            2024.05.08.593163"
COMMENT     Sequence Label: pME_Cp_0_7-8_003_Amp/ColEI_mScarlet-I
FEATURES             Location/Qualifiers
     source          1..2883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             163..174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      885..956
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      972..999
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1120..1911,1912..1980))
                     /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     1743..1762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1981..2085)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2188..2776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2677..2696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacatgagac cagagaaagt gaaacgtgat ttcatgcgtc attttgaaca ttttgtaaat
       61 cttatttaat aatgtgtgcg gcaattcaca tttaatttat gaatgttttc ttaacatcgc
      121 ggcaactcaa gaaacggcag gttcggatct tagctactag agaaagagga gaaatactag
      181 atgcgtaaag gcgaagcagt gatcaaagaa tttatgcgct tcaaagttca catggaaggt
      241 tctatgaacg gccacgaatt tgaaattgaa ggtgaaggcg aaggtcgtcc atacgaaggt
      301 actcaaaccg caaaactgaa agttaccaaa ggtggtccac tgccattttc ttgggatatt
      361 ctgtctccac aatttatgta cggttctcgt gcatttatca aacacccagc agatattcca
      421 gactactaca aacaatcttt tccggaaggt ttcaaatggg aacgtgttat gaattttgaa
      481 gatggtggtg cagttacggt tacccaagat acctctctgg aagatggtac tctgatctac
      541 aaagttaaac tgcgtggtac taactttcca ccagatggtc cagttatgca gaaaaaaacc
      601 atgggttggg aagcatctac cgaacgtctg tacccagaag atggcgttct gaaaggtgat
      661 atcaaaatgg cactgcgtct gaaagatggc ggtcgttacc tggcagattt caaaaccacc
      721 tacaaagcga aaaaaccagt tcaaatgcca ggtgcataca acgttgatcg taaactggat
      781 attaccagcc acaacgaaga ttacaccgtt gttgaacaat acgaacgttc tgaaggccgt
      841 cactctaccg gtggtatgga tgaactgtac aaataaccag gcatcaaata aaacgaaagg
      901 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
      961 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggtct caagactgcg
     1021 gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1081 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1141 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1201 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1261 ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1321 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1381 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1441 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1501 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1561 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1621 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1681 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1741 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1801 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1861 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1921 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1981 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2041 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2101 agtgccacct gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2821 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc
     2881 cgc
//
