LOCUS       sequence-12801-e        7501 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 7501)
  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 7501)
  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-che-DAS+4
FEATURES             Location/Qualifiers
     source          1..7501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(21..37)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(30..52)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    596..730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(633..652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     687..706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      complement(783..810)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(900..917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1071..1659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1151..1170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5868..5884
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5869..5885
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6098..6120
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6117..6133
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6117..6133
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6153..6173
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    6177..6201
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6693..6717)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6735..7442
                     /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(6880..6898)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(join(7494..7501,1..13))
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ttatacatag ttgataattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
       61 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggggattg
      241 gtggcgacga ctcctggagc ccgtcagtat cggcggaatt ccagctgagc gccggtcgct
      301 accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg
      361 tatttcgcgt aaggaaatcc attatgtact atttaaaaaa cacaaacttt tggatgttcg
      421 gtttattctt tttcttttac ttttttatca tgggagccta cttcccgttt ttcccgattt
      481 ggctacatga catcaaccat atcagcaaaa gtgatacggg tattattttt gccgctattt
      541 ctctgttctc gctattattc caaccgctgt ttggtctgct ttctgacaaa ctcggaactt
      601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      721 tgtctggatc ctctagagtc gacctgcagg tttaaacgcg gccgcaaaca gtattaaacg
      781 caaaaaggcc atccgtcagg atggccttct gcttagctaa tcaactagtt ctagctgcat
      841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1501 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccggacg tggccgacca
     1921 gcccgtcatc gtcaacgcct gaccgcggtg cggacaggcc gtgtcgcgac cggccgtgcg
     1981 gaattaagcc ggcccgtacc ctgtgaatag aggtccgctg tgacacaaga atccctgtta
     2041 cttctcgacc gtattgattc ggatgattcc tacgcgagcc tgcggaacga ccaggaattc
     2101 tgggagccgc tggcccgccg agccctggag gagctcgggc tgccggtgcc gccggtgctg
     2161 cgggtgcccg gcgagagcac caaccccgta ctggtcggcg agcccgaccc ggtgatcaag
     2221 ctgttcggcg agcactggtg cggtccggag agcctcgcgt cggagtcgga ggcgtacgcg
     2281 gtcctggcgg acgccccggt gccggtgccc cgcctcctcg gccgcggcga gctgcggccc
     2341 ggcaccggag cctggccgtg gccctacctg gtgatgagcc ggatgaccgg caccacctgg
     2401 cggtccgcga tggacggcac gaccgaccgg aacgcgctgc tcgccctggc ccgcgaactc
     2461 ggccgggtgc tcggccggct gcacagggtg ccgctgaccg ggaacaccgt gctcaccccc
     2521 cattccgagg tcttcccgga actgctgcgg gaacgccgcg cggcgaccgt cgaggaccac
     2581 cgcgggtggg gctacctctc gccccggctg ctggaccgcc tggaggactg gctgccggac
     2641 gtggacacgc tgctggccgg ccgcgaaccc cggttcgtcc acggcgacct gcacgggacc
     2701 aacatcttcg tggacctggc cgcgaccgag gtcaccggga tcgtcgactt caccgacgtc
     2761 tatgcgggag actcccgcta cagcctggtg caactgcatc tcaacgcctt ccggggcgac
     2821 cgcgagatcc tggccgcgct gctcgacggg gcgcagtgga agcggaccga ggacttcgcc
     2881 cgcgaactgc tcgccttcac cttcctgcac gacttcgagg tgttcgagga gaccccgctg
     2941 gatctctccg gcttcaccga tccggaggaa ctggcgcagt tcctctgggg gccgccggac
     3001 accgcccccg gcgcctgacg ccccgggccg cccggcgccg cccccggccc ccggcggccg
     3061 gccgcccgga gccccgcccg cgctcgggag ccccgggccc gcgccgaagc ccgctgctgc
     3121 gagcccggag cgggccggcc gacggcggtg cgggcccggc ggcggacgct cagcagcggc
     3181 gggcgtgaaa ggccctggca tcctcgatca tctcctccag ggtggtcggc cgagcttcca
     3241 tcccagctcg gcaaggatca acgaggacag tcgcacgacg aagttcttct ggatcgcgcc
     3301 cgtgctggaa gcactcaacc tcgaagcgtg tggttgcgga gccatctagc aaccacacga
     3361 aacatgcgca acgaaccgcg caacgaacaa cgcctagaac tggccctaga tgagctgact
     3421 cgtatcgttg gtaaacctag tttgaccagc atgttttaac tacgttcggt gagctgtcaa
     3481 cggggcctgt aacggcacaa cgaaccgtgc aacgagagtg gccacggatg ccaccacagg
     3541 cactacaacg gagttcgcca cgtacatcac cacaaccacc gattctggcg gtgagctccc
     3601 cgatattcag cggaaatggc ttggtatcga ccaagattcg tagaaccccg tctcgtctgg
     3661 ctggtattca aaacgggcgc aacgaaacac gcaacgagac aggcatggcc caaaccagaa
     3721 aactagcgtc taccaggact tttacctgtc cgacccgttg caacggaacc ccccacggaa
     3781 cccccgcgac acccgctccc caattgcgtt agaacagcgg tggattgtcg gcttcgttgt
     3841 gggccttttg agccgcttcc tgttctgccg cacgctcttt cctcgcccga tagccgagtc
     3901 gcttaacggt gtccagatgc agcccgaaat gtttggccgt ttgcggccaa gagtggccct
     3961 cgtcgtcgtg ataggcgcgg atgcgttcgc ggcgtgcagc ctgctcggcg agccactcgc
     4021 tgcgttcctg cgccacgagc cggacgacgt ggcgttcgga tagtccggtg attcgagcgc
     4081 cttcggcggc ggtcacgcgc cgctttttgc ggacagtcgg ctgccggttg tagccgtcgc
     4141 tgtagccgtc gctgtagccg tcgctcatag caatgcctcc atggctgacg cggactttgc
     4201 gcgccgcgca actgtgctcg ccgccgtgcg cgctgctgcg cccttccgcg agatggccga
     4261 ctggcgcgca ctgagtgtgg cctcgtagac cacgatcccg tccgcccaaa tgcgcgactt
     4321 ggttgtgatc caacgccaaa tgctgttggc gatggcgcgg acctcgctgt ccggtagcgg
     4381 tccgggacac acgtcgttgc acgggaattc ggcgtttcgc gcgtggcact cggcatagat
     4441 cgcgcggccg agtccgtcca cgttccgggt cggcaggtag atccgcatga gggcgggacg
     4501 ataggcccac aacctgacgg aatcgaacag tgcgcaattc cgccctagcg gcgtcggagc
     4561 cgctttgtac gtggtctgct gacgccagcg cggcggtggc atgttcgcgc cgagctcggc
     4621 ctcgatgtgg ctgagtgtgt agagatctga gtggagccat tccgtttccc aggcgatgtg
     4681 gccggggttt ttggtcatga ggcctgagta actgcggtcg ccgtcgacgg cgcgccgaag
     4741 gccttcggcg cacgccgcca tgtatgcgag cggcttacgc cgcgcgtatt cggtgcgtgg
     4801 aacaggggcg ttgagtgccc acactgcgtg tgcgtggccg ttggcgcgat tgcccacgat
     4861 cgcgttgggc agcggatggg acccccgggc gctgagcgct cggagcgctg cgtctggatg
     4921 gtctacgtcc acgaccagca ggtttgccag cgctgttggg ttcgcctcga tgtaccggcg
     4981 gcctagggcc gacgcgcggc tttggcggta gatcccctcg agcagatcgt cgcttgccag
     5041 cggccagtac ggcagccaga gctgctcaaa ttcgtcggcg acgtggctca cgcttggtag
     5101 tagaccacga ttaatcaccg gtgtatggtc cgacacgagc tccaagtcag atatttcgct
     5161 gaggggccac cccacaactg cacactcccc cgctctcccg tcgagccctg gtggtggaac
     5221 accagcgaca gccgagcacc cccaaccacc tgtaccaacc aggaggaaca catgcgtcgt
     5281 ttcgaggacg tttccgggcc gctgagagcc gctgtggcgg ccgtacacgc cgccttagac
     5341 ccgttagacc ccctgccgcc tgaatgcgcg ggtacgagcc acacagcgcc cgaacttacg
     5401 gagctggtgg gctcacctgg ctttatggcg tacgaatcgg ctgtgtgcga cctgttgggc
     5461 gaggtgaggt acgcgctact cacgctggca agggcgacac agccgcccca ccgagcccgc
     5521 acggccgcgc gcggtgtcaa caaccgggtg agtcgtgcac accagcaggt gttcgaggct
     5581 tggctcgaag tgcaggacat cgtggcgaac gccgcccgat gagccgcgcc ttacgctggc
     5641 tgccagccgt tcgcgggctg gttggtgcag cgcgtcgagc ggttagaggc cctgcggtgt
     5701 tccaccaccg caggcctcgc cctttttaag gctgaatttg cttgtctccg aatccaactg
     5761 gcttgtccaa gggtgtatct acgcttaatc caaagttcaa acgaggggat tacacatgac
     5821 caacttcgat aacgttctcg gctcgatgat ccccggcggg ccccccctcg aggtcgacgg
     5881 tatcgattta attaatatgc atcaattgat ttaaatacta gtgttaacta tttaattggg
     5941 gaccctagag gtcccctttt ttattttaaa aattttttcn aaggggacct ctagggtccc
     6001 cagctggcta gcgagtcatg aggtnaccat gttctttcct gcgttatccc ctgattctgt
     6061 ggataaccgt attaccgcct ttgagtgagc tgataccagc ggataacaat ttcacacagg
     6121 aaacagctat gaccatgatt acgccaagct atcaactttg tatagaaaag ttgaattcaa
     6181 gtttgtacaa aaaagcaggc ttattggatc acgccgcgag cgtgtcggtg ttcgaagacc
     6241 tgggaatcca gcaggcgccc aacagcaccg tctgctccgt cttctccaga aaggcgacta
     6301 cgcggtcctc ctcgtcggtg ggtacgcagc cgaatgcccg cgggtcgccc acccgcacca
     6361 gttgcagcgt gacatcggct cgattgcttc ggtggaagtc cagcagttgg gccagatccg
     6421 cgagggcccc ccgtgaacat cgtcgtggta accagcgtgt cggctttggc tgtggccgtg
     6481 gtgcattcgg tggcgtttgc gatcaagggg cgtcacggcg aggacctggt cgaccgctgg
     6541 aaccgtgagg gcatcggtgc acaccgcggg atcagaccgt cgacgaacca tttgaatccg
     6601 accggtacct caacgagttg acggccgatc acgtccaggt tatgttcgat gacgatcacc
     6661 gtattgccct tgtcgaccag gccgttgatc cgacccagct ttcttgtaca aagtgggcat
     6721 gcaggaggtt aataatggtg agcaagggcg aggaggacaa catggccatc atcaaggagt
     6781 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6841 gcgagggcga gggccgcccg tacgagggca cccagaccgc caagctgaag gtgaccaagg
     6901 gtggcccgct gccgttcgcc tgggacatcc tgtcgccaca gttcatgtac ggctcgaagg
     6961 cctacgtgaa gcacccggcc gacatcccgg actacttgaa gctgtccttc ccggagggct
     7021 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7081 cgtccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccgt
     7141 cggacggccc ggtcatgcag aagaagacca tgggctggga ggcctcgagc gagcggatgt
     7201 acccggagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7261 gccactacga cgcggaggtc aagaccacct acaaggccaa gaagccggtg cagctgccgg
     7321 gcgcctacaa cgtcaacatc aagttggaca tcacctcgca caacgaggac tacaccatcg
     7381 tggaacagta cgaacgcgcc gagggccgcc actcgaccgg cggtatggac gagctgtaca
     7441 aggccgcgaa cgatgagaat tacagcgaaa actacgccga cgcgtcgtag tgacaacttt
     7501 a
//
