LOCUS       sequence-469960-        2904 bp    DNA     circular SYN 24-APR-2025
DEFINITION  Ampicillin/ColEI backbone with sfGFP placeholder for lvl1 assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2904)
  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 2904)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 24, 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_004_Amp/ColEI_sfGFP
FEATURES             Location/Qualifiers
     source          1..2904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             163..174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             181..894
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      906..977
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      993..1020
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1141..1932,1933..2001))
                     /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     1764..1783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2002..2106)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2209..2797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2698..2717
                     /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 gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
      241 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
      301 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggttccttg gccgactctg
      361 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
      421 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
      481 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
      541 aaccgcattg agctgaaagg cattgacttt aaagaggacg gcaatatcct gggccataag
      601 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
      661 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
      721 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
      781 ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
      841 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgacca
      901 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      961 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     1021 tttataggtc tcaagactgc ggccgcgatt atcaaaaagg atcttcacct agatcctttt
     1081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1261 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
     1321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2101 tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt
     2161 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2221 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2281 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2341 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2401 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2461 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2521 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2581 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2641 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2701 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2761 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2821 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2881 ttctgtggat aaccgtgcgg ccgc
//
