LOCUS       sequence-14203-e        4777 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 4777)
  AUTHORS   Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller 
            JA, Somia NV, Bogdanove AJ, Voytas DF
  TITLE     Efficient design and assembly of custom TALEN and other TAL 
            effector-based constructs for DNA targeting.
  JOURNAL   Nucleic Acids Res. 2011 Jul;39(12):e82. doi: 10.1093/nar/gkr218
  PUBMED    21493687
REFERENCE   2  (bases 1 to 4777)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2011-07"; volume: "39(12):e82. doi"; pages: " 
            10.1093/nar/gkr21"
COMMENT     Sequence Label: pTAL1
FEATURES             Location/Qualifiers
     source          1..4777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(927..1334)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1199..1215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1225..1243
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1291..1311)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1291..1309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1330..1346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1343..1365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1354..1370
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1378..1384,1385..1402,1403..1408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1423..1444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(2285..2384)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2402..2420)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2425..2441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(2603..3394,3395..3463))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3226..3245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3591..4179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4080..4099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4333..4350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4509..4536)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4628..4714)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgaattc gcccttttgc atgtaaatag gaggtgcacc atggatccca
      181 ttcgtccgcg caggccaagt cctgcccgcg agcttctgcc cggaccccaa ccggataggg
      241 ttcagccgac tgcagatcgt ggggtgtctg cgcctgctgg cagccctctg gatggcttgc
      301 ccgctcggcg gacggtgtcc cggacccggc tgccatctcc ccctgcgccc tcacctgcgt
      361 tctcggcggg cagcttcagc gatctgctcc gtccgttcga tccgtcgctt cttgatacat
      421 cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc ccagcagagt
      481 gggatgaggc gcaatcggct ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg
      541 ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac
      601 cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac agtcagcagc
      661 agcaagagaa gatcaaaccg aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg
      721 tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag
      781 ggaccgtcgc tgtcacgtat cagcacataa tcacggcgtt gccagaggcg acacacgaag
      841 acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg
      901 atgcggggga gttgagaggt ccgccgttac agttggacac aggccaactt gtgaagattg
      961 caaaacgtgg cggcgtgacc gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg
     1021 gtgcccccct ggagacgggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     1081 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     1141 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     1201 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg
     1261 ccccccctcg aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     1321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1441 attaattgcg ttgcgctcac tgcccgcttt ccaccggtcg tctccaacga ccacctcgtc
     1501 gccttggcct gcctcggcgg acgtcctgcc atggatgcag tgaaaaaggg attgccgcac
     1561 gcgccggaat tgatcagaag agtcaatcgc cgtattggcg aacgcacgtc ccatcgcgtt
     1621 gccgactacg cgcaagtggt tcgcgtgctg gagtttttcc agtgccactc ccacccagcg
     1681 tacgcatttg atgaggccat gacgcagttc gggatgagca ggaacgggtt ggtacagctc
     1741 tttcgcagag tgggcgtcac cgaactcgaa gcccgcggtg gaacgctccc cccagcctcg
     1801 cagcgttggg accgtatcct ccaggcatca gggatgaaaa gggccaaacc gtcccctact
     1861 tcagctcaaa caccggatca ggcgtctttg catgcattcg ccgattcgct ggagcgtgac
     1921 cttgatgcgc ccagcccaat gcacgaggga gatcagacgc gggcaagcag ccgtaaacgg
     1981 tcccgatcgg atcgtgctgt caccggcccc tccgcacagc aggctgtcga ggtgcgcgtt
     2041 cccgaacagc gcgatgcgct gcatttgccc ctcagctgga gggtaaaacg cccgcgtacc
     2101 aggatctggg gcggcctccc ggatcctggt acgcccatgg ctgccgacct ggcagcgtcc
     2161 agcaccgtga tgtgggaaca agatgcggac cccttcgcag gggcagcgga tgatttcccg
     2221 gcattcaacg aagaggaact cgcatggttg atggagctat tgcctcagtg aaagggcgaa
     2281 ttcgacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2341 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2401 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2461 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaga ttatcaaaaa
     2521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3481 attattgaag catttatcag ggttattgtc tcatgaccaa aatcccttaa cgtgagttac
     3541 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3661 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3721 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3781 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3841 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3901 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3961 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4021 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4081 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4141 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4201 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4261 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4321 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4381 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4441 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     4501 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     4561 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     4621 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     4681 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4741 ttaacgctag catggatgtt ttcccagtca cgacgtt
//
