LOCUS       sequence-433363-        5337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy glmS-nadE plasmid containing mCherry and no antibiotic 
            resistance gene (ARG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5337)
  AUTHORS   Amrofell MB, Rengarajan S, Vo ST, Ramirez Tovar ES, LoBello L, 
            Dantas G, Moon TS
  TITLE     Engineering E. coli strains using antibiotic-resistance-gene-free 
            plasmids.
  JOURNAL   Cell Rep Methods. 2023 Dec 18;3(12):100669. doi: 
            10.1016/j.crmeth.2023.100669. Epub 2023 Dec 11.
  PUBMED    38086386
REFERENCE   2  (bases 1 to 5337)
  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: "Cell Rep 
            Methods."; date: "2023-12-18"; volume: "3(12):100669. doi"; pages: "
            10.1016/j.crmeth.2023.100669. Epub 2023 Dec 11"
COMMENT     Sequence Label: pMBA329
FEATURES             Location/Qualifiers
     source          1..5337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1738..1755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1989..2008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2668..2702
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             2723..2731
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2737..3446
                     /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
                     EGRHSTGGMESCTS"
     primer_bind     complement(2882..2900)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3150..3169
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3585..3603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4197..4231
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
ORIGIN
        1 gctggcctgc gtctgtcgaa agagctgggt taccttggtt cactggcaat ctgtaacgtt
       61 ccgggttctt ctctggtgcg cgaatccgat ctggcgctaa tgaccaacgc gggtacagaa
      121 atcggcgtgg catccactaa agcattcacc actcagttaa ctgtgctgtt gatgctggtg
      181 gcgaagctgt ctcgcctgaa aggtctggat gcctccattg aacatgacat cgtgcatggt
      241 ctgcaggcgc tgccgagccg tattgagcag atgctgtctc aggacaaacg cattgaagcg
      301 ctggcagaag atttctctga caaacatcac gcgctgttcc tgggccgtgg cgatcagtac
      361 ccaatcgcgc tggaaggcgc attgaagttg aaagagatct cttacattca cgctgaagcc
      421 tacgctgctg gcgaactgaa acacggtccg ctggcgctaa ttgatgccga tatgccggtt
      481 attgttgttg caccgaacaa cgaattgctg gaaaaactga aatccaacat tgaagaagtt
      541 cgcgcgcgtg gcggtcagtt gtatgtcttc gccgatcagg atgcgggttt tgtaagtagc
      601 gataacatgc acatcatcga gatgccgcat gtggaagagg tgattgcacc gatcttctac
      661 accgttccgc tgcagctgct ggcttaccat gtcgcgctga tcaaaggcac cgacgttgac
      721 cagccgcgta acctggcaaa atcggttacg gttgagtaag aacgatacct aatagaaaat
      781 aggaattgac accaatgaca ttgcaacaac aaataataaa ggcgctgggc gcaaaaccgc
      841 agattaatgc tgaagaggaa attcgtcgta gtatcgattt tctgaaaagc tacctgcaaa
      901 cttatccgtt cattaaatca ctggtgctcg ggatcagcgg cggtcaggac tccacgcttg
      961 ccggtaagct gtgccagatg gcgattaatg agctgcgcca ggaaaccggc aacgaatcac
     1021 tgcaatttat tgccgtacgc ctgccctatg gtgtccaggc cgacgaacaa gattgccagg
     1081 atgccatcgc ctttattcaa ccggatcgcg tattaaccgt taatatcaag ggcgcggtac
     1141 tcgccagcga gcaggcattg cgggaagcag gcattgaact gagcgatttt gtccgtggca
     1201 atgaaaaagc gcgtgagcgg atgaaagcac aatatagcat tgcgggtatg accagcggtg
     1261 tcgtggtggg caccgatcat gctgcagaag ctattaccgg attcttcact aaatatggtg
     1321 acggcggtac ggacattaat ccgctgtatc gtctcaacaa acgtcagggt aaacagttac
     1381 tggcggcatt aggttgcccg gaacaccttt ataagaaagc gccaacggcc gatcttgaag
     1441 atgatcgccc ttctctgccg gatgaagtgg cactcggcgt gacctatgac aatatcgacg
     1501 actatctgga agggaaaaac gtacctgaac aggtcgccag aacaatagag aactggtatc
     1561 tgaaaaccga acataaacgc cgtccgccaa ttaccgtttt cgatgatttc tggaaaaagt
     1621 aaagccgagc aagacaatgg taagtagtaa tagctcctgc ccccggaaga tcattccggg
     1681 ggctttttta ttcccactgt gaactggatc cttactcgag tctagactgc aggcttcctc
     1741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct
     1921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2401 agttggtagc tcttgatccg ccaaacaaac caccgctggt agcggtggtt tttttgtttg
     2461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2641 tatatatgag taaacttggt ctgacagttg acagctagct cagtcctagg tataatgcta
     2701 gcttcttaat tacgtaggac cataaggagg tttcctatgg tgagcaaggg cgaggaggat
     2761 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2821 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2881 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     2941 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3001 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3061 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3121 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3181 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3241 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3301 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3361 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3421 ggcggcatgg agagctgtac aagtaagcgg aagagttaag accattcctc ttacgtgccg
     3481 atcaacgtct cattttcgcc agatatcggg ttccgcgcac atttccccga aaagtgccac
     3541 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     3601 ggcatcaccc taggtgtaaa acgacggcca gactcggtac caaattccag aaaagagacg
     3661 ctgaaaagcg tcttttttcg ttttggtcct tattaatttt taaagtatgg gcttaaccag
     3721 attagatccc gtagatgtcc atccccgccg aaacgtttgg tggcgggacc agtgacgaag
     3781 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     3841 ctccagcgaa agcggtcctc tccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     3901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     3961 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc gggcttatcg
     4021 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
     4081 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
     4141 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgaccctga
     4201 cagctagctc agtcctaggt ataatgctag cgtcaattta gtagtagacc acaaggaggt
     4261 tgcttaatgt gtggaattgt tggcgcgatc gcgcaacgtg atgtagcaga aatccttctt
     4321 gaaggtttac gtcgtctgga ataccgcgga tatgactctg ccggtctggc cgttgttgat
     4381 gcagaaggtc atatgacccg cctgcgtcgc ctcggtaaag tccagatgct ggcacaggca
     4441 gcggaagaac atcctctgca tggcggcact ggtattgctc acactcgctg ggcgacccac
     4501 ggtgaacctt cagaagtgaa tgcgcatccg catgtttctg aacacattgt ggtggtgcat
     4561 aacggcatca tcgaaaacca tgaaccgctg cgtgaagagc taaaagcgcg tggctatacc
     4621 ttcgtttctg aaaccgacac cgaagtgatt gcccatctgg tgaactggga gctgaaacaa
     4681 ggcgggactc tgcgtgaggc cgttctgcgt gctatcccgc agctgcgtgg tgcgtacggt
     4741 acagtgatca tggactcccg tcacccggat accctgctgg cggcacgttc tggtagtccg
     4801 ctggtgattg gcctggggat gggcgaaaac tttatcgctt ctgaccagct ggcgctgttg
     4861 ccggtgaccc gtcgctttat cttccttgaa gagggcgata ttgcggaaat cactcgccgt
     4921 tcggtaaaca tcttcgataa aactggcgcg gaagtaaaac gtcaggatat cgaatccaat
     4981 ctgcaatatg acgcgggcga taaaggcatt taccgtcact acatgcagaa agagatctac
     5041 gaacagccga acgcgatcaa aaacaccctt accggacgca tcagccacgg tcaggttgat
     5101 ttaagcgagc tgggaccgaa cgccgacgaa ctgctgtcga aggttgagca tattcagatc
     5161 ctcgcctgtg gtacttctta taactccggt atggtttccc gctactggtt tgaatcgcta
     5221 gcaggtattc cgtgcgacgt cgaaatcgcc tctgaattcc gctatcgcaa atctgccgtg
     5281 cgtcgtaaca gcctgatgat caccttgtca cagtctggcg aaaccgcgga taccctg
//
