LOCUS       sequence-12794-e        6573 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 6573)
  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 6573)
  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-GCT1
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      144..153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(675..677,678..680,681..1391)
                     /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(708..736)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1331..1351
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      complement(1473..1500)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1590..1607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1761..2349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1841..1860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6558..6573,1)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     join(6559..6573,1..2)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgatttaatt aatatgcatc aattgattta aatactagtg ttaactattt aattggggac
       61 cctagaggtc ccctttttta ttttaaaaat tttttcnaag gggacctcta gggtccccag
      121 ctggctagcg agtcatgagg tnaccatggt ggccatctag aaatattgga tcacgccgcg
      181 agcgtgtcgg tgttcgaaga cctgggaatc cagcaggcgc ccaacagcac cgtctgctcc
      241 gtcttctcca gaaaggcgac tacgcggtcc tcctcgtcgg tgggtacgca gccgaatgcc
      301 cgcgggtcgc ccacccgcac cagttgcagc gtgacatcgg ctcgattgct tcggtggaag
      361 tccagcagtt gggccagatc cgcgagggcc ccccgtgaac atcgtcgtgg taaccagcgt
      421 gtcggctttg gctgtggccg tggtgcattc ggtggcgttt gcgatcaagg ggcgtcacgg
      481 cgaggacctg gtcgaccgct ggaaccgtga gggcatcggt gcacaccgcg ggatcagacc
      541 gtcgacgaac catttgaatc cgaccggtac ctcaacgagt tgacggccga tcacgtccag
      601 gttatgttcg atgacgatca ccgtattgcc cttgtcgacc aggccgttga tccgcatgcg
      661 aaggagatat acatatggct agcaaaggag aagaactttt cactggagtt gtcccaattc
      721 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
      781 gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga aaactacctg
      841 ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttacc
      901 cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
      961 aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg
     1021 aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa
     1081 acattctcgg acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag
     1141 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggct
     1201 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     1261 taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc
     1321 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
     1381 agctctacaa aggcgccgca aacgacgaaa actacgcttt agcagcttaa tgactgcagg
     1441 tttaaacgcg gccgcaaaca gtattaaacg caaaaaggcc atccgtcagg atggccttct
     1501 gcttagctaa tcaactagtt ctagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1561 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2161 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     2221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2581 gttgcctgac tccccggacg tggccgacca gcccgtcatc gtcaacgcct gaccgcggtg
     2641 cggacaggcc gtgtcgcgac cggccgtgcg gaattaagcc ggcccgtacc ctgtgaatag
     2701 aggtccgctg tgacacaaga atccctgtta cttctcgacc gtattgattc ggatgattcc
     2761 tacgcgagcc tgcggaacga ccaggaattc tgggagccgc tggcccgccg agccctggag
     2821 gagctcgggc tgccggtgcc gccggtgctg cgggtgcccg gcgagagcac caaccccgta
     2881 ctggtcggcg agcccgaccc ggtgatcaag ctgttcggcg agcactggtg cggtccggag
     2941 agcctcgcgt cggagtcgga ggcgtacgcg gtcctggcgg acgccccggt gccggtgccc
     3001 cgcctcctcg gccgcggcga gctgcggccc ggcaccggag cctggccgtg gccctacctg
     3061 gtgatgagcc ggatgaccgg caccacctgg cggtccgcga tggacggcac gaccgaccgg
     3121 aacgcgctgc tcgccctggc ccgcgaactc ggccgggtgc tcggccggct gcacagggtg
     3181 ccgctgaccg ggaacaccgt gctcaccccc cattccgagg tcttcccgga actgctgcgg
     3241 gaacgccgcg cggcgaccgt cgaggaccac cgcgggtggg gctacctctc gccccggctg
     3301 ctggaccgcc tggaggactg gctgccggac gtggacacgc tgctggccgg ccgcgaaccc
     3361 cggttcgtcc acggcgacct gcacgggacc aacatcttcg tggacctggc cgcgaccgag
     3421 gtcaccggga tcgtcgactt caccgacgtc tatgcgggag actcccgcta cagcctggtg
     3481 caactgcatc tcaacgcctt ccggggcgac cgcgagatcc tggccgcgct gctcgacggg
     3541 gcgcagtgga agcggaccga ggacttcgcc cgcgaactgc tcgccttcac cttcctgcac
     3601 gacttcgagg tgttcgagga gaccccgctg gatctctccg gcttcaccga tccggaggaa
     3661 ctggcgcagt tcctctgggg gccgccggac accgcccccg gcgcctgacg ccccgggccg
     3721 cccggcgccg cccccggccc ccggcggccg gccgcccgga gccccgcccg cgctcgggag
     3781 ccccgggccc gcgccgaagc ccgctgctgc gagcccggag cgggccggcc gacggcggtg
     3841 cgggcccggc ggcggacgct cagcagcggc gggcgtgaaa ggccctggca tcctcgatca
     3901 tctcctccag ggtggtcggc cgagcttcca tcccagctcg gcaaggatca acgaggacag
     3961 tcgcacgacg aagttcttct ggatcgcgcc cgtgctggaa gcactcaacc tcgaagcgtg
     4021 tggttgcgga gccatctagc aaccacacga aacatgcgca acgaaccgcg caacgaacaa
     4081 cgcctagaac tggccctaga tgagctgact cgtatcgttg gtaaacctag tttgaccagc
     4141 atgttttaac tacgttcggt gagctgtcaa cggggcctgt aacggcacaa cgaaccgtgc
     4201 aacgagagtg gccacggatg ccaccacagg cactacaacg gagttcgcca cgtacatcac
     4261 cacaaccacc gattctggcg gtgagctccc cgatattcag cggaaatggc ttggtatcga
     4321 ccaagattcg tagaaccccg tctcgtctgg ctggtattca aaacgggcgc aacgaaacac
     4381 gcaacgagac aggcatggcc caaaccagaa aactagcgtc taccaggact tttacctgtc
     4441 cgacccgttg caacggaacc ccccacggaa cccccgcgac acccgctccc caattgcgtt
     4501 agaacagcgg tggattgtcg gcttcgttgt gggccttttg agccgcttcc tgttctgccg
     4561 cacgctcttt cctcgcccga tagccgagtc gcttaacggt gtccagatgc agcccgaaat
     4621 gtttggccgt ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg atgcgttcgc
     4681 ggcgtgcagc ctgctcggcg agccactcgc tgcgttcctg cgccacgagc cggacgacgt
     4741 ggcgttcgga tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc cgctttttgc
     4801 ggacagtcgg ctgccggttg tagccgtcgc tgtagccgtc gctgtagccg tcgctcatag
     4861 caatgcctcc atggctgacg cggactttgc gcgccgcgca actgtgctcg ccgccgtgcg
     4921 cgctgctgcg cccttccgcg agatggccga ctggcgcgca ctgagtgtgg cctcgtagac
     4981 cacgatcccg tccgcccaaa tgcgcgactt ggttgtgatc caacgccaaa tgctgttggc
     5041 gatggcgcgg acctcgctgt ccggtagcgg tccgggacac acgtcgttgc acgggaattc
     5101 ggcgtttcgc gcgtggcact cggcatagat cgcgcggccg agtccgtcca cgttccgggt
     5161 cggcaggtag atccgcatga gggcgggacg ataggcccac aacctgacgg aatcgaacag
     5221 tgcgcaattc cgccctagcg gcgtcggagc cgctttgtac gtggtctgct gacgccagcg
     5281 cggcggtggc atgttcgcgc cgagctcggc ctcgatgtgg ctgagtgtgt agagatctga
     5341 gtggagccat tccgtttccc aggcgatgtg gccggggttt ttggtcatga ggcctgagta
     5401 actgcggtcg ccgtcgacgg cgcgccgaag gccttcggcg cacgccgcca tgtatgcgag
     5461 cggcttacgc cgcgcgtatt cggtgcgtgg aacaggggcg ttgagtgccc acactgcgtg
     5521 tgcgtggccg ttggcgcgat tgcccacgat cgcgttgggc agcggatggg acccccgggc
     5581 gctgagcgct cggagcgctg cgtctggatg gtctacgtcc acgaccagca ggtttgccag
     5641 cgctgttggg ttcgcctcga tgtaccggcg gcctagggcc gacgcgcggc tttggcggta
     5701 gatcccctcg agcagatcgt cgcttgccag cggccagtac ggcagccaga gctgctcaaa
     5761 ttcgtcggcg acgtggctca cgcttggtag tagaccacga ttaatcaccg gtgtatggtc
     5821 cgacacgagc tccaagtcag atatttcgct gaggggccac cccacaactg cacactcccc
     5881 cgctctcccg tcgagccctg gtggtggaac accagcgaca gccgagcacc cccaaccacc
     5941 tgtaccaacc aggaggaaca catgcgtcgt ttcgaggacg tttccgggcc gctgagagcc
     6001 gctgtggcgg ccgtacacgc cgccttagac ccgttagacc ccctgccgcc tgaatgcgcg
     6061 ggtacgagcc acacagcgcc cgaacttacg gagctggtgg gctcacctgg ctttatggcg
     6121 tacgaatcgg ctgtgtgcga cctgttgggc gaggtgaggt acgcgctact cacgctggca
     6181 agggcgacac agccgcccca ccgagcccgc acggccgcgc gcggtgtcaa caaccgggtg
     6241 agtcgtgcac accagcaggt gttcgaggct tggctcgaag tgcaggacat cgtggcgaac
     6301 gccgcccgat gagccgcgcc ttacgctggc tgccagccgt tcgcgggctg gttggtgcag
     6361 cgcgtcgagc ggttagaggc cctgcggtgt tccaccaccg caggcctcgc cctttttaag
     6421 gctgaatttg cttgtctccg aatccaactg gcttgtccaa gggtgtatct acgcttaatc
     6481 caaagttcaa acgaggggat tacacatgac caacttcgat aacgttctcg gctcgatgat
     6541 ccccggcggg ccccccctcg aggtcgacgg tat
//
