LOCUS       sequence-12798-e        6589 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 6589)
  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 6589)
  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: pTE-0X38-GCT6
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      149..158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(680..682,683..685,686..1396)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(713..741)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1336..1356
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      complement(1494..1521)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1611..1628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1782..2370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1862..1881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6579..6589,1..6)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     join(6580..6589,1..7)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtatcgatt taattaatat gcatcaattg atttaaatac tagtgttaac tatttaattg
       61 gggaccctag aggtcccctt ttttatttta aaaatttttt cnaaggggac ctctagggtc
      121 cccagctggc tagcgagtca tgaggtnacc atggtggcca tctagaaata ttggatcacg
      181 ccgcgagcgt gtcggtgttc gaagacctgg gaatccagca ggcgcccaac agcaccgtct
      241 gctccgtctt ctccagaaag gcgactacgc ggtcctcctc gtcggtgggt acgcagccga
      301 atgcccgcgg gtcgcccacc cgcaccagtt gcagcgtgac atcggctcga ttgcttcggt
      361 ggaagtccag cagttgggcc agatccgcga gggccccccg tgaacatcgt cgtggtaacc
      421 agcgtgtcgg ctttggctgt ggccgtggtg cattcggtgg cgtttgcgat caaggggcgt
      481 cacggcgagg acctggtcga ccgctggaac cgtgagggca tcggtgcaca ccgcgggatc
      541 agaccgtcga cgaaccattt gaatccgacc ggtacctcaa cgagttgacg gccgatcacg
      601 tccaggttat gttcgatgac gatcaccgta ttgcccttgt cgaccaggcc gttgatccgc
      661 atgcgaagga gatatacata tggctagcaa aggagaagaa cttttcactg gagttgtccc
      721 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      781 tgaaggtgat gctacatacg gaaagcttac ccttaaattt atttgcacta ctggaaaact
      841 acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat gcttttcccg
      901 ttacccggat cacatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
      961 acaggaacgc actatatctt tcaaagatga cgggaactac aagacgcgtg ctgaagtcaa
     1021 gtttgaaggt gatacccttg ttaatcgtat cgagttaaaa ggtattgatt ttaaagaaga
     1081 tggaaacatt ctcggacaca aactcgagta caactataac tcacacaatg tatacatcac
     1141 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca acattgaaga
     1201 tggctccgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     1261 ccttttacca gacaaccatt acctgtcgac acaatctgcc ctttcgaaag atcccaacga
     1321 aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta cacatggcat
     1381 ggatgagctc tacaaaggat ccgccgcaaa cgacgagaac tactccgaga actacgcgga
     1441 cgccagctaa tgagctgcag gtttaaacgc ggccgcaaac agtattaaac gcaaaaaggc
     1501 catccgtcag gatggccttc tgcttagcta atcaactagt tctagctgca ttaatgaatc
     1561 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1621 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1681 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2221 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2581 tgtctatttc gttcatccat agttgcctga ctccccggac gtggccgacc agcccgtcat
     2641 cgtcaacgcc tgaccgcggt gcggacaggc cgtgtcgcga ccggccgtgc ggaattaagc
     2701 cggcccgtac cctgtgaata gaggtccgct gtgacacaag aatccctgtt acttctcgac
     2761 cgtattgatt cggatgattc ctacgcgagc ctgcggaacg accaggaatt ctgggagccg
     2821 ctggcccgcc gagccctgga ggagctcggg ctgccggtgc cgccggtgct gcgggtgccc
     2881 ggcgagagca ccaaccccgt actggtcggc gagcccgacc cggtgatcaa gctgttcggc
     2941 gagcactggt gcggtccgga gagcctcgcg tcggagtcgg aggcgtacgc ggtcctggcg
     3001 gacgccccgg tgccggtgcc ccgcctcctc ggccgcggcg agctgcggcc cggcaccgga
     3061 gcctggccgt ggccctacct ggtgatgagc cggatgaccg gcaccacctg gcggtccgcg
     3121 atggacggca cgaccgaccg gaacgcgctg ctcgccctgg cccgcgaact cggccgggtg
     3181 ctcggccggc tgcacagggt gccgctgacc gggaacaccg tgctcacccc ccattccgag
     3241 gtcttcccgg aactgctgcg ggaacgccgc gcggcgaccg tcgaggacca ccgcgggtgg
     3301 ggctacctct cgccccggct gctggaccgc ctggaggact ggctgccgga cgtggacacg
     3361 ctgctggccg gccgcgaacc ccggttcgtc cacggcgacc tgcacgggac caacatcttc
     3421 gtggacctgg ccgcgaccga ggtcaccggg atcgtcgact tcaccgacgt ctatgcggga
     3481 gactcccgct acagcctggt gcaactgcat ctcaacgcct tccggggcga ccgcgagatc
     3541 ctggccgcgc tgctcgacgg ggcgcagtgg aagcggaccg aggacttcgc ccgcgaactg
     3601 ctcgccttca ccttcctgca cgacttcgag gtgttcgagg agaccccgct ggatctctcc
     3661 ggcttcaccg atccggagga actggcgcag ttcctctggg ggccgccgga caccgccccc
     3721 ggcgcctgac gccccgggcc gcccggcgcc gcccccggcc cccggcggcc ggccgcccgg
     3781 agccccgccc gcgctcggga gccccgggcc cgcgccgaag cccgctgctg cgagcccgga
     3841 gcgggccggc cgacggcggt gcgggcccgg cggcggacgc tcagcagcgg cgggcgtgaa
     3901 aggccctggc atcctcgatc atctcctcca gggtggtcgg ccgagcttcc atcccagctc
     3961 ggcaaggatc aacgaggaca gtcgcacgac gaagttcttc tggatcgcgc ccgtgctgga
     4021 agcactcaac ctcgaagcgt gtggttgcgg agccatctag caaccacacg aaacatgcgc
     4081 aacgaaccgc gcaacgaaca acgcctagaa ctggccctag atgagctgac tcgtatcgtt
     4141 ggtaaaccta gtttgaccag catgttttaa ctacgttcgg tgagctgtca acggggcctg
     4201 taacggcaca acgaaccgtg caacgagagt ggccacggat gccaccacag gcactacaac
     4261 ggagttcgcc acgtacatca ccacaaccac cgattctggc ggtgagctcc ccgatattca
     4321 gcggaaatgg cttggtatcg accaagattc gtagaacccc gtctcgtctg gctggtattc
     4381 aaaacgggcg caacgaaaca cgcaacgaga caggcatggc ccaaaccaga aaactagcgt
     4441 ctaccaggac ttttacctgt ccgacccgtt gcaacggaac cccccacgga acccccgcga
     4501 cacccgctcc ccaattgcgt tagaacagcg gtggattgtc ggcttcgttg tgggcctttt
     4561 gagccgcttc ctgttctgcc gcacgctctt tcctcgcccg atagccgagt cgcttaacgg
     4621 tgtccagatg cagcccgaaa tgtttggccg tttgcggcca agagtggccc tcgtcgtcgt
     4681 gataggcgcg gatgcgttcg cggcgtgcag cctgctcggc gagccactcg ctgcgttcct
     4741 gcgccacgag ccggacgacg tggcgttcgg atagtccggt gattcgagcg ccttcggcgg
     4801 cggtcacgcg ccgctttttg cggacagtcg gctgccggtt gtagccgtcg ctgtagccgt
     4861 cgctgtagcc gtcgctcata gcaatgcctc catggctgac gcggactttg cgcgccgcgc
     4921 aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg actggcgcgc
     4981 actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact tggttgtgat
     5041 ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg gtccgggaca
     5101 cacgtcgttg cacgggaatt cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc
     5161 gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggac gataggccca
     5221 caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta
     5281 cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg
     5341 gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt ggccggggtt
     5401 tttggtcatg aggcctgagt aactgcggtc gccgtcgacg gcgcgccgaa ggccttcggc
     5461 gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc
     5521 gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg
     5581 cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc
     5641 cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc
     5701 cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca gcggccagta
     5761 cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttggta gtagaccacg
     5821 attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc tgaggggcca
     5881 ccccacaact gcacactccc ccgctctccc gtcgagccct ggtggtggaa caccagcgac
     5941 agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg tttcgaggac
     6001 gtttccgggc cgctgagagc cgctgtggcg gccgtacacg ccgccttaga cccgttagac
     6061 cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc ccgaacttac ggagctggtg
     6121 ggctcacctg gctttatggc gtacgaatcg gctgtgtgcg acctgttggg cgaggtgagg
     6181 tacgcgctac tcacgctggc aagggcgaca cagccgcccc accgagcccg cacggccgcg
     6241 cgcggtgtca acaaccgggt gagtcgtgca caccagcagg tgttcgaggc ttggctcgaa
     6301 gtgcaggaca tcgtggcgaa cgccgcccga tgagccgcgc cttacgctgg ctgccagccg
     6361 ttcgcgggct ggttggtgca gcgcgtcgag cggttagagg ccctgcggtg ttccaccacc
     6421 gcaggcctcg ccctttttaa ggctgaattt gcttgtctcc gaatccaact ggcttgtcca
     6481 agggtgtatc tacgcttaat ccaaagttca aacgagggga ttacacatga ccaacttcga
     6541 taacgttctc ggctcgatga tccccggcgg gccccccctc gaggtcgac
//
