LOCUS       sequence-193817-        3578 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses mCherry fluorescent protein in E. coli. The expression is 
            controlled by a pGlnA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3578)
  AUTHORS   Guo S, Yeung E, Murray RM
  TITLE     Implementation and System Identification of a Phosphorylation-Based 
            Insulator in a Cell-Free Transcription-Translation System
  JOURNAL   bioRxiv 122606
REFERENCE   2  (bases 1 to 3578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            122606"
COMMENT     Sequence Label: pSG10-pGlnA-mCherry
FEATURES             Location/Qualifiers
     source          1..3578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             345..1055
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(490..508)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     758..777
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     rep_origin      1346..1890
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1791..1810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2007..2024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(2257..3048,3049..3117))
                     /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     2880..2899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3118..3222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3290..3308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atcatcttat taatcagata aaatatttct agagcatcct ccgcaaacaa gtattgcaga
       61 gtccctttgt gatcgctttc acggagcata aaaagggtta tccaaaggtc attgcaccaa
      121 catggtgctt aatgtttcca ttgaagcact atattggtgc aacattcaca tcgtggtgca
      181 gcccttttgc acgatggtgc gcatgataac gccttttagg ggcaatttaa aagttggcac
      241 agatttcgct ttatcttttt tacggcgaca cggccaaaat aattgcagat ttcgttacca
      301 cgacgaccta actttcaatt ctaataagga ggaagacttc aaccatggtg agcaagggcg
      361 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      421 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      481 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      541 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      601 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      661 acggcggcgt ggtgaccgtg acccaggact cctccttgca ggacggcgag ttcatctaca
      721 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      781 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
      841 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
      901 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
      961 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     1021 actccaccgg cggcatggac gagctgtaca agtaataatc tcgagcaaag cccgccgaaa
     1081 ggcgggcttt tctgtgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     1141 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     1201 tcgtaggaca ggtgccggca tgataagctg tcaaacatga gaattacaac ttatatcgta
     1261 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt
     1321 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     1381 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     1441 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     1501 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     1561 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     1621 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     1681 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     1741 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag gggaaacgcc
     1801 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     1861 tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc
     1921 tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg
     1981 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc
     2041 tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac
     2101 ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagggtcat
     2161 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2221 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2281 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2341 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2401 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2461 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2521 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2581 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2641 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2701 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2761 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2821 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2881 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2941 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3001 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3061 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3121 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3181 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3241 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3301 cacgaggccc tttcgtcttc aagaattctg gcgaatcctc tgaccagcca gaaaacgacc
     3361 tttctgtggt gaaaccggat gctgcaattc agagcggcag caagtggggg acagcagaag
     3421 acctgaccgc cgcagagtgg atgtttgaca tggtgaagac tatcgcacca tcagccagaa
     3481 aaccgaattt tgctgggtgg gctaacgata tccgcctgat gcgtgaacgt gacggacgta
     3541 accaccgcga catgtgtgtg ctgttccgct gggcatgc
//
