LOCUS       sequence-13865-e        6345 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 6345)
  AUTHORS   Takaki K, Davis JM, Winglee K, Ramakrishnan L
  TITLE     Evaluation of the pathogenesis and treatment of Mycobacterium 
            marinum infection in zebrafish.
  JOURNAL   Nat Protoc. 2013 Jun;8(6):1114-24. doi: 10.1038/nprot.2013.068. Epub
            2013 May 16.
  PUBMED    23680983
REFERENCE   2  (bases 1 to 6345)
  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: 
            "10.1038/nprot.2013.068"; journalName: "Nat Protoc"; date: 
            "2013-06"; volume: "8"; issue: "6"; pages: "1114-24"
COMMENT     Sequence Label: pTEC18
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..49)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             join(104..106,107..109,110..823)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(149..170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(952..971)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1036..1055)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3146..3734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3635..3654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5797..5813
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5797..5813
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc
       61 ggtaccagat ctttaaatct agatttaaga aggagatata catatggtga gcaagggcga
      121 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      181 caagttcagc gtgaggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
      241 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag
      301 ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      361 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcac
      421 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      481 gaagggcgtc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
      541 caacagccac aacatctata tcatggccgt caagcagaag aacggcatca aggtgaactt
      601 caagatccgc cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa
      661 cacccccatc ggcgacggcc ccgtgctgct gcccgacagc cactacctga gcacccagtc
      721 cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttccgcac
      781 cgccgccggg atcactctcg gcatggacga gctgtacaag taagctagcc acctgacgtc
      841 gggggggggg ggaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
      901 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
      961 ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg
     1021 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1081 tctatcgctt gtatgggaag ccccatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1141 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1201 ctcttccgac catcaagcat tttatccgta ctcctgatga ctagagcatg agattatcaa
     1261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1381 cgatctgtct atttcgttca tccatagttg cctgactccc cgacgtggcc gaccagcccg
     1441 tcatcgtcaa cgcctgatcc gcggtgcgga caggccgtgt cgcgaccggc cgtgcggaat
     1501 taagccggcc cgtaccctgt gaatagaggt ccgctgtgac acaagaatcc ctgttacttc
     1561 tcgaccgtat tgattcggat gattcctacg cgagcctgcg gaacgaccag gaattctggg
     1621 agccgctggc ccgccgagcc ctggaggagc tcgggctgcc ggtgccgccg gtgctgcggg
     1681 tgcccggcga gagcaccaac cccgtactgg tcggcgagcc cggcccggtg atcaagctgt
     1741 tcggcgagca ctggtgcggt ccggagagcc tcgcgtcgga gtcggaggcg tacgcggtcc
     1801 tggcggacgc cccggtgccg gtgccccgcc tcctcggccg cggcgagctg cggcccggca
     1861 ccggagcctg gccgtggccc tacctggtga tgagccggat gaccggcacc acctggcggt
     1921 ccgcgatgga cggcacgacc gaccggaacg cgctgctcgc cctggcccgc gaactcggcc
     1981 gggtgctcgg ccggctgcac agggtgccgc tgaccgggaa caccgtgctc accccccatt
     2041 ccgaggtctt cccggaactg ctgcgggaac gccgcgcggc gaccgtcgag gaccaccgcg
     2101 ggtggggcta cctctcgccc cggctgctgg accgcctgga ggactggctg ccggacgtgg
     2161 acacgctgct ggccggccgc gaaccccggt tcgtccacgg cgacctgcac gggaccaaca
     2221 tcttcgtgga cctggccgcg accgaggtca ccgggatcgt cgacttcacc gacgtctatg
     2281 cgggagactc ccgctacagc ctggtgcaac tgcatctcaa cgccttccgg ggcgaccgcg
     2341 agatcctggc cgcgctgctc gacggggcgc agtggaagcg gaccgaggac ttcgcccgcg
     2401 aactgctcgc cttcaccttc ctgcacgact tcgaggtgtt cgaggagacc ccgctggatc
     2461 tctccggctt caccgatccg gaggaactgg cgcagttcct ctgggggccg ccggacaccg
     2521 cccccggcgc ctgacgcccc ctctagtaat cttttgccat tctcaccgga ttcagtcgtc
     2581 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     2641 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     2701 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2761 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     2821 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt
     2881 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg
     2941 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag
     3001 ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt
     3061 atgagtcagc aacaccttct tcacgaggca gacctcacta gttccactga gcgtcagacc
     3121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     3541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcaac gcgtgagccc
     3901 accagctccg taagttcggg tgctgtgtgg ctcgtacccg cgcattcagg cggcaggggg
     3961 tctaacgggt ctaaggcggc gtgtacggcc gccacagcgg ctcttagcgg cccggaaacg
     4021 tcctcgaaac gacgcatgtg ttcctcctgg ttggtacagg tggttggggg tgctcggctg
     4081 tcgctggtgt ttcatcatca gggctcgacg ggagagcggg ggagtgtgca gttgtggggt
     4141 ggcccctcag cgaaatatct gacttggagc tcgtgtcgga ccatacaccg gtgattaatc
     4201 gtggtttatt atcaagcgtg agccacgtcg ccgacgaatt tgagcagctc tggctgccgt
     4261 actggtccct ggcaagcgac gatctgctcg aggggatcta ccgccaaagc cgcgcgtcgg
     4321 ccctaggccg ccggtacatc gaggcgaacc caacagcgct ggcaaacctg ctggtcgtgg
     4381 acgtagacca tccagacgca gcgctccgag cgctcagcgc ccgggggtcc catccgctgc
     4441 ccaacgcgat cgtgggcaat cgcgccaacg gccacgcaca cgcagtgtgg gcactcaacg
     4501 cccctgttcc acgcaccgaa tacgcgcggc gtaagccgct cgcatacatg gcggcgtgcg
     4561 ccgaaggcct tcggcgcgcc gtcgatggcg accgcagtta ctcaggcctc atgaccaaaa
     4621 accccggcca catcgcctgg gaaacggaat ggctccactc agatctctac acactcagcc
     4681 acatcgaggc cgagctcggc gcgaacatgc caccgccgcg ctggcgtcag cagaccacgt
     4741 acaaagcggc tccgacgccg ctagggcgga attgcgcact gttcgattcc gtcaggttgt
     4801 gggcctatct tcccgccctc atgcggatct acctgccgac ccggaacgtg gacggactcg
     4861 gccgcgcgat ctatgccgag tgccacgcgc gaaacgccga atttccgtgc aacgacgtgt
     4921 gtcccggacc gctaccggac agcgaggtcc gcgccatcgc caacagcatt tggcgttgga
     4981 tcacaaccaa gtcgcgcatt tgggcggacg ggatcgtggt ctacgaggcc acactcagtg
     5041 cgcgccatgc ggccatctcg cggaagggcg cagcagcgcg cacggcggcg agcacagttg
     5101 cgcggcgcgc aaagtccgcg tcagccatgg aggcattgct atgagcgacg gctacagcga
     5161 cggctacagc gacggctaca actggcagcc gactgtccgc aaaaagcggc gcgtgaccgc
     5221 cgccgaaggc gctcgaatca ccggactatc cgaacgccac gtcgtccggc tcgtggcgca
     5281 ggaacgcagc gagtggttcg ccgagcaggc tgcacgccgc gaacgcatcc gcgcctatca
     5341 cgacgacgag ggccactctt ggccgcaaac ggccaaacat ttcgggctgc atctggacac
     5401 cgttaagcga ctcggctatc gggcgaggaa agagcgtgcg gcagaacagg aagcggctca
     5461 aaaggcccac aacgaagccg acaatccacc gctgttctaa cgcaattggg gagcgggtgt
     5521 cgcgggggtt ccgtgggggg ttccgttgca acgggtcgga caggtaaaag tcctggtaga
     5581 cgctagtttt ctggtttggg ccatgcctgt ctcgttgcgt gtttcgttgc gtccgttttg
     5641 aataccagcc agacgagacg gggttctacg aatcttggtc gataccaagc catttccgct
     5701 gaatatcgtg gagctcaccg ccagaatcgg tggttgtggt gatgtacgtg gcgaactccg
     5761 ttgtagtgct tgtggtggca tccgtggcgc ggccgctcta gaactagtgg atctgacccg
     5821 ctccacaacc gctgagcagg gtggcagtca gagcgaccag gccgaccgat acgaggcgga
     5881 ttttgaacag gttcactcga agcctcccgc tgggttcgga gcatcgtcga tcacatgacg
     5941 gcgtgctgac accgattcga ctatgcacag tagtaggtag ccgccgcgga cggtagcgca
     6001 ggcggcctcg gcgccgtcgt ggcgtcgcga tggttggcgc cagacccgcc tgatcggcgt
     6061 cccgaccagg gccgccgggg tggtgagcct cggcttggtc cagccgtagt gagacgggat
     6121 gtcttcccgg ttgtcgggtt tgggtgggcg gtcacatgtt cgctggctgt gggcgtgtcg
     6181 ggggtcgtcg aagcggccgc gaacagggct ggacgatgca cgtcttgcac ctcctgtcaa
     6241 cccccaatgt tgagccgcga tgcccctgac ctgcaccgtt ggaccgcagt cgttaggggg
     6301 gcgtgttagc atggagtgac gaaaggggcc taaatcagat gatcc
//
