LOCUS       sequence-200305-        3579 bp    DNA     circular SYN 15-DEC-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 3579)
  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 3579)
  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 
            122606"
COMMENT     Sequence Label: pSG10-pGlnA-mCherry
FEATURES             Location/Qualifiers
     source          1..3579
                     /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..1891
                     /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     1792..1811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2008..2025
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(2258..3049,3050..3118))
                     /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     2881..2900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3119..3223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3291..3309
                     /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 gaattgcaac 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 ggggaaacgc
     1801 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     1861 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     1921 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     1981 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     2041 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     2101 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc gctagggtca
     2161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2521 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2581 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2641 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2701 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2761 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2821 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2881 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2941 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3001 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3061 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3121 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3181 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3241 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3301 tcacgaggcc ctttcgtctt caagaattct ggcgaatcct ctgaccagcc agaaaacgac
     3361 ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca gcaagtgggg gacagcagaa
     3421 gacctgaccg ccgcagagtg gatgtttgac atggtgaaga ctatcgcacc atcagccaga
     3481 aaaccgaatt ttgctgggtg ggctaacgat atccgcctga tgcgtgaacg tgacggacgt
     3541 aaccaccgcg acatgtgtgt gctgttccgc tgggcatgc
//
