LOCUS       sequence-12799-e        7486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7486)
  AUTHORS   Kim JH, Wei JR, Wallach JB, Robbins RS, Rubin EJ, Schnappinger D
  TITLE     Protein inactivation in mycobacteria by controlled proteolysis and 
            its application to deplete the beta subunit of RNA polymerase.
  JOURNAL   Nucleic Acids Res. 2011 Mar;39(6):2210-20. doi: 10.1093/nar/gkq1149
  PUBMED    21075796
REFERENCE   2  (bases 1 to 7486)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2011-03"; volume: "39"; issue: "6"; 
            pages: "2210-20"
COMMENT     Sequence Label: pGMEH-P38-mRFP
FEATURES             Location/Qualifiers
     source          1..7486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..19
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4..20
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     233..255
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     252..268
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     252..268
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    288..308
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    312..336
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(828..852)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             870..1580
                     /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(1015..1033)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(1614..1634)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(1642..1659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1642..1658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1651..1673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2217..2351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2254..2273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2308..2327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      complement(2404..2431)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2521..2538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2692..3280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2772..2791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctcgaggtc gacggtatcg atttaattaa tatgcatcaa ttgatttaaa tactagtgtt
       61 aactatttaa ttggggaccc tagaggtccc cttttttatt ttaaaaattt tttcnaaggg
      121 gacctctagg gtccccagct ggctagcgag tcatgaggtn accatgttct ttcctgcgtt
      181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccagcggata
      241 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctatcaa ctttgtatag
      301 aaaagttgaa ttcaagtttg tacaaaaaag caggcttatt ggatcacgcc gcgagcgtgt
      361 cggtgttcga agacctggga atccagcagg cgcccaacag caccgtctgc tccgtcttct
      421 ccagaaaggc gactacgcgg tcctcctcgt cggtgggtac gcagccgaat gcccgcgggt
      481 cgcccacccg caccagttgc agcgtgacat cggctcgatt gcttcggtgg aagtccagca
      541 gttgggccag atccgcgagg gccccccgtg aacatcgtcg tggtaaccag cgtgtcggct
      601 ttggctgtgg ccgtggtgca ttcggtggcg tttgcgatca aggggcgtca cggcgaggac
      661 ctggtcgacc gctggaaccg tgagggcatc ggtgcacacc gcgggatcag accgtcgacg
      721 aaccatttga atccgaccgg tacctcaacg agttgacggc cgatcacgtc caggttatgt
      781 tcgatgacga tcaccgtatt gcccttgtcg accaggccgt tgatccgacc cagctttctt
      841 gtacaaagtg ggcatgcagg aggttaataa tggtgagcaa gggcgaggag gacaacatgg
      901 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
      961 agttcgagat cgagggcgag ggcgagggcc gcccgtacga gggcacccag accgccaagc
     1021 tgaaggtgac caagggtggc ccgctgccgt tcgcctggga catcctgtcg ccacagttca
     1081 tgtacggctc gaaggcctac gtgaagcacc cggccgacat cccggactac ttgaagctgt
     1141 ccttcccgga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1201 ccgtgaccca ggactcgtcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1261 gcaccaactt cccgtcggac ggcccggtca tgcagaagaa gaccatgggc tgggaggcct
     1321 cgagcgagcg gatgtacccg gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1381 agctgaagga cggcggccac tacgacgcgg aggtcaagac cacctacaag gccaagaagc
     1441 cggtgcagct gccgggcgcc tacaacgtca acatcaagtt ggacatcacc tcgcacaacg
     1501 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcg accggcggta
     1561 tggacgagct gtacaagtag tgacccgggc tgcaggaatt cgatatcaag cttcaacttt
     1621 attatacata gttgataatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggggatt
     1861 ggtggcgacg actcctggag cccgtcagta tcggcggaat tccagctgag cgccggtcgc
     1921 taccattacc agttggtctg gtgtcaaaaa taataataac cgggcaggcc atgtctgccc
     1981 gtatttcgcg taaggaaatc cattatgtac tatttaaaaa acacaaactt ttggatgttc
     2041 ggtttattct ttttctttta cttttttatc atgggagcct acttcccgtt tttcccgatt
     2101 tggctacatg acatcaacca tatcagcaaa agtgatacgg gtattatttt tgccgctatt
     2161 tctctgttct cgctattatt ccaaccgctg tttggtctgc tttctgacaa actcggaact
     2221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2341 atgtctggat cctctagagt cgacctgcag gtttaaacgc ggccgcaaac agtattaaac
     2401 gcaaaaaggc catccgtcag gatggccttc tgcttagcta atcaactagt tctagctgca
     2461 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2521 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3121 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccggac gtggccgacc
     3541 agcccgtcat cgtcaacgcc tgaccgcggt gcggacaggc cgtgtcgcga ccggccgtgc
     3601 ggaattaagc cggcccgtac cctgtgaata gaggtccgct gtgacacaag aatccctgtt
     3661 acttctcgac cgtattgatt cggatgattc ctacgcgagc ctgcggaacg accaggaatt
     3721 ctgggagccg ctggcccgcc gagccctgga ggagctcggg ctgccggtgc cgccggtgct
     3781 gcgggtgccc ggcgagagca ccaaccccgt actggtcggc gagcccgacc cggtgatcaa
     3841 gctgttcggc gagcactggt gcggtccgga gagcctcgcg tcggagtcgg aggcgtacgc
     3901 ggtcctggcg gacgccccgg tgccggtgcc ccgcctcctc ggccgcggcg agctgcggcc
     3961 cggcaccgga gcctggccgt ggccctacct ggtgatgagc cggatgaccg gcaccacctg
     4021 gcggtccgcg atggacggca cgaccgaccg gaacgcgctg ctcgccctgg cccgcgaact
     4081 cggccgggtg ctcggccggc tgcacagggt gccgctgacc gggaacaccg tgctcacccc
     4141 ccattccgag gtcttcccgg aactgctgcg ggaacgccgc gcggcgaccg tcgaggacca
     4201 ccgcgggtgg ggctacctct cgccccggct gctggaccgc ctggaggact ggctgccgga
     4261 cgtggacacg ctgctggccg gccgcgaacc ccggttcgtc cacggcgacc tgcacgggac
     4321 caacatcttc gtggacctgg ccgcgaccga ggtcaccggg atcgtcgact tcaccgacgt
     4381 ctatgcggga gactcccgct acagcctggt gcaactgcat ctcaacgcct tccggggcga
     4441 ccgcgagatc ctggccgcgc tgctcgacgg ggcgcagtgg aagcggaccg aggacttcgc
     4501 ccgcgaactg ctcgccttca ccttcctgca cgacttcgag gtgttcgagg agaccccgct
     4561 ggatctctcc ggcttcaccg atccggagga actggcgcag ttcctctggg ggccgccgga
     4621 caccgccccc ggcgcctgac gccccgggcc gcccggcgcc gcccccggcc cccggcggcc
     4681 ggccgcccgg agccccgccc gcgctcggga gccccgggcc cgcgccgaag cccgctgctg
     4741 cgagcccgga gcgggccggc cgacggcggt gcgggcccgg cggcggacgc tcagcagcgg
     4801 cgggcgtgaa aggccctggc atcctcgatc atctcctcca gggtggtcgg ccgagcttcc
     4861 atcccagctc ggcaaggatc aacgaggaca gtcgcacgac gaagttcttc tggatcgcgc
     4921 ccgtgctgga agcactcaac ctcgaagcgt gtggttgcgg agccatctag caaccacacg
     4981 aaacatgcgc aacgaaccgc gcaacgaaca acgcctagaa ctggccctag atgagctgac
     5041 tcgtatcgtt ggtaaaccta gtttgaccag catgttttaa ctacgttcgg tgagctgtca
     5101 acggggcctg taacggcaca acgaaccgtg caacgagagt ggccacggat gccaccacag
     5161 gcactacaac ggagttcgcc acgtacatca ccacaaccac cgattctggc ggtgagctcc
     5221 ccgatattca gcggaaatgg cttggtatcg accaagattc gtagaacccc gtctcgtctg
     5281 gctggtattc aaaacgggcg caacgaaaca cgcaacgaga caggcatggc ccaaaccaga
     5341 aaactagcgt ctaccaggac ttttacctgt ccgacccgtt gcaacggaac cccccacgga
     5401 acccccgcga cacccgctcc ccaattgcgt tagaacagcg gtggattgtc ggcttcgttg
     5461 tgggcctttt gagccgcttc ctgttctgcc gcacgctctt tcctcgcccg atagccgagt
     5521 cgcttaacgg tgtccagatg cagcccgaaa tgtttggccg tttgcggcca agagtggccc
     5581 tcgtcgtcgt gataggcgcg gatgcgttcg cggcgtgcag cctgctcggc gagccactcg
     5641 ctgcgttcct gcgccacgag ccggacgacg tggcgttcgg atagtccggt gattcgagcg
     5701 ccttcggcgg cggtcacgcg ccgctttttg cggacagtcg gctgccggtt gtagccgtcg
     5761 ctgtagccgt cgctgtagcc gtcgctcata gcaatgcctc catggctgac gcggactttg
     5821 cgcgccgcgc aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg
     5881 actggcgcgc actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact
     5941 tggttgtgat ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg
     6001 gtccgggaca cacgtcgttg cacgggaatt cggcgtttcg cgcgtggcac tcggcataga
     6061 tcgcgcggcc gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggac
     6121 gataggccca caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag
     6181 ccgctttgta cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg
     6241 cctcgatgtg gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt
     6301 ggccggggtt tttggtcatg aggcctgagt aactgcggtc gccgtcgacg gcgcgccgaa
     6361 ggccttcggc gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg
     6421 gaacaggggc gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga
     6481 tcgcgttggg cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat
     6541 ggtctacgtc cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc
     6601 ggcctagggc cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca
     6661 gcggccagta cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttggta
     6721 gtagaccacg attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc
     6781 tgaggggcca ccccacaact gcacactccc ccgctctccc gtcgagccct ggtggtggaa
     6841 caccagcgac agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg
     6901 tttcgaggac gtttccgggc cgctgagagc cgctgtggcg gccgtacacg ccgccttaga
     6961 cccgttagac cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc ccgaacttac
     7021 ggagctggtg ggctcacctg gctttatggc gtacgaatcg gctgtgtgcg acctgttggg
     7081 cgaggtgagg tacgcgctac tcacgctggc aagggcgaca cagccgcccc accgagcccg
     7141 cacggccgcg cgcggtgtca acaaccgggt gagtcgtgca caccagcagg tgttcgaggc
     7201 ttggctcgaa gtgcaggaca tcgtggcgaa cgccgcccga tgagccgcgc cttacgctgg
     7261 ctgccagccg ttcgcgggct ggttggtgca gcgcgtcgag cggttagagg ccctgcggtg
     7321 ttccaccacc gcaggcctcg ccctttttaa ggctgaattt gcttgtctcc gaatccaact
     7381 ggcttgtcca agggtgtatc tacgcttaat ccaaagttca aacgagggga ttacacatga
     7441 ccaacttcga taacgttctc ggctcgatga tccccggcgg gccccc
//
