LOCUS       sequence-433362-        4748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium 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 4748)
  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 4748)
  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: pMBA328
FEATURES             Location/Qualifiers
     source          1..4748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(840..934)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        961..995
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             1016..1024
                     /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             1030..1740
                     /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(1175..1193)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1443..1462
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1793..1864
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1880..1907
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1935..1952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2069..2614)
                     /direction=LEFT
                     /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     complement(2149..2168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2720..2754
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
ORIGIN
        1 gctacctgca aacttatccg ttcattaaat cactggtgct cgggatcagc ggcggtcagg
       61 actccacgct tgccggtaag ctgtgccaga tggcgattaa tgagctgcgc caggaaaccg
      121 gcaacgaatc actgcaattt attgccgtac gcctgcccta tggtgtccag gccgacgaac
      181 aagattgcca ggatgccatc gcctttattc aaccggatcg cgtattaacc gttaatatca
      241 agggcgcggt actcgccagc gagcaggcat tgcgggaagc aggcattgaa ctgagcgatt
      301 ttgtccgtgg caatgaaaaa gcgcgtgagc ggatgaaagc acaatatagc attgcgggta
      361 tgaccagcgg tgtcgtggtg ggcaccgatc atgctgcaga agctattacc ggattcttca
      421 ctaaatatgg tgacggcggt acggacatta atccgctgta tcgtctcaac aaacgtcagg
      481 gtaaacagtt actggcggca ttaggttgcc cggaacacct ttataagaaa gcgccaacgg
      541 ccgatcttga agatgatcgc ccttctctgc cggatgaagt ggcactcggc gtgacctatg
      601 acaatatcga cgactatctg gaagggaaaa acgtacctga acaggtcgcc agaacaatag
      661 agaactggta tctgaaaacc gaacataaac gccgtccgcc aattaccgtt ttcgatgatt
      721 tctggaaaaa gtaaagcctc aaagtaatca agccctggta actgccaggg cttttttatt
      781 tcatctttga atcaacatat taaaattcat cgaatggcat ccttgctaac agtgcttgta
      841 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
      901 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa ccccagagtc
      961 ttgacagcta gctcagtcct aggtataatg ctagcttctt aattacgtag gaccataagg
     1021 aggtttccta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1081 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1141 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1201 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1261 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     1321 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1381 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1441 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1501 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     1561 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1621 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     1681 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     1741 gcggaagagt taagaccatt ggatccaaac tcgagtaagg atctccaggc atcaaataaa
     1801 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1861 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacctaggg
     1921 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     1981 aaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     2041 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     2101 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     2161 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     2221 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     2281 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     2341 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     2401 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     2461 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     2521 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     2581 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     2641 atatttctag atttcagtgc aatttatctc ttcaaatgta gcagaggcta gtgtagatcg
     2701 ctactagagc gcactgaccc tgacagctag ctcagtccta ggtataatgc tagcgtcaat
     2761 ttagtagtag accacaagga ggttgcttaa tgtgtggaat tgttggcgcg atcgcgcaac
     2821 gtgatgtagc agaaatcctt cttgaaggtt tacgtcgtct ggaataccgc ggatatgact
     2881 ctgccggtct ggccgttgtt gatgcagaag gtcatatgac ccgcctgcgt cgcctcggta
     2941 aagtccagat gctggcacag gcagcggaag aacatcctct gcatggcggc actggtattg
     3001 ctcacactcg ctggacgacc cacggtgaac cttcagaagt gaatgcgcat ccgcatgttt
     3061 ctgaacacat tgtggtggtg cataacggca tcatcgaaaa ccatgaaccg ctgcgtgaag
     3121 agctaaaagc gcgtggctat accttcgttt ctgaaaccga caccgaagtg attgcccatc
     3181 tggtgaactg ggagctgaaa caaggcggga ctctgcgtga ggccgttctg cgtgctatcc
     3241 cgcagctgcg tggtgcgtac ggtacagtga tcatggactc ccgtcacccg gataccctgc
     3301 tggcggcacg ttctggtagt ccgctggtga ttggcctggg gatgggcgaa aactttatcg
     3361 cttctgacca gctggcgctg ttgccggtga cccgtcgctt tatcttcctt gaagagggcg
     3421 atattgcgga aatcactcgc cgttcggtaa acatcttcga taaaactggc gcggaagtaa
     3481 aacgtcagga tatcgaatcc aatctgcaat atgacgcggg cgataaaggc atttaccgtc
     3541 actacatgca gaaagagatc tacgaacagc cgaacgcgat caaaaacacc cttaccggac
     3601 gcatcagcca cggtcaggtt gatttaagcg agctgggacc gaacgccgac gaactgctgt
     3661 cgaaggttga gcatattcag atcctcgcct gtggtacttc ttataactcc ggtatggttt
     3721 cccgctactg gtttgaatcg ctagcaggta ttccgtgcga cgtcgaaatc gcctctgaat
     3781 tccgctatca caaatctgcc gtgcgtcgta acagcctgat gatcaccttg tcacagtctg
     3841 gcgaaaccgc ggataccctg gctggcctgc gtctgtcgaa agagctgggt taccttggtt
     3901 cactggcaat ctgtaacgtt ccgggttctt ctctggtgcg cgaatccgat ctggcgctaa
     3961 tgaccaacgc gggtacagaa atcggcgtgg catccactaa agcattcacc actcagttaa
     4021 ctgtgctgtt gatgctggtg gcgaagctgt ctcgcctgaa aggtctggat gcctccattg
     4081 aacatgacat cgtgcatggt ctgcaggcgc tgccgagccg tattgagcag atgctgtctc
     4141 aggacaaacg cattgaagcg ctggcagaag atttctctga caaacatcac gcgctgttcc
     4201 tgggccgtgg cgatcagtac ccaatcgcgc tggaaggcgc attgaagttg aaagagatct
     4261 cttacattca cgctgaagcc tacgctgctg gcgaactgaa acacggtccg ctggcgctaa
     4321 ttgatgccga tatgccggtt attgttgttg caccgaacaa cgaattgctg gaaaaactga
     4381 aatccaacat tgaagaagtt cgcgcgcgtg gcggtcagtt gtatgtcttc gccgatcagg
     4441 atgcgggttt tgtaagtagc gataacatgc acatcatcga gatgccgcat gtggaagagg
     4501 tgattgcacc gatcttctac accgttccgc tgcagctgct ggcttaccat gtcgcgctga
     4561 tcaaaggcac cgacgttgac cagccgcgta acctggcaaa atcggttacg gttgagtaag
     4621 aacgatacct aatagaaaat aggaattgac accaatgaca ttgcaacaac aaataataaa
     4681 ggcgctgggc gcaaaaccgc agattaatgc tgaagaggaa attcgtcgta gtatcgattt
     4741 tctgaaaa
//
