LOCUS       sequence-62295-e        6090 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 6090)
  AUTHORS   Zhang Y, Zhang F, Li X, Baller JA, Qi Y, Starker CG, Bogdanove AJ, 
            Voytas DF
  TITLE     TALENs enable efficient plant genome engineering.
  JOURNAL   Plant Physiol. 2012 Nov 2.
  PUBMED    23124327
REFERENCE   2  (bases 1 to 6090)
  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: "Plant 
            Physiol. 2012 Nov 2."
COMMENT     Sequence Label: pZHY051
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1028..1373
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1316..1336
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    1432..1556
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1581..1586,1587..1604,1605..1611)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1665..2345
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(1731..1750)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2665..2970
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2849..2868
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3011..3135)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3201..3453
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3471..3487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3471..3487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3484..3506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3495..3511
                     /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(3519..3525,3526..3543,3544..3549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3564..3585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3702..3719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3953..3972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4632..5423,5424..5492))
                     /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     5255..5274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5493..5597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5665..5683
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5721..5743)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5843..5862
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6056..6078
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6070..6087
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6071..6087
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttgcat gcctgcagtg cagcgtgacc aaatctcaga attccaatcc cacaaaaatc
       61 tgagcttaac agcacagttg ctcctctcag agcagaatcg ggtattcaac accctcatat
      121 caactactac gttgtgtata acggtccaca tgccggtata tacgatgact ggggttgtac
      181 aaaggcggca acaaacggcg ttcccggagt tgcacacaag aaatttgcca ctattacaga
      241 ggcaagagca gcagctgacg cgtacacaac aagtcagcaa acagacaggt tgaacttcat
      301 ccccaaagga gaagctcaac tcaagcccaa gagctttgct aaggccctaa caagcccacc
      361 aaagcaaaaa gcccactggc tcacgctagg aaccaaaagg cccagcagtg atccagcccc
      421 aaaagagatc tcctttgccc cggagattac aatggacgat ttcctctatc tttacgatct
      481 aggaaggaag ttcgaaggtg aaggtgacga cactatgttc accactgata atgagaaggt
      541 tagcctcttc aatttcagaa agaatgctga cccacagatg gttagagagg cctacgcagc
      601 aggtctcatc aagacgatct acccgagtaa caatctccag gagatcaaat accttcccaa
      661 gaaggttaaa gatgcagtca aaagattcag gactaattgc atcaagaaca cagagaaaga
      721 catatttctc aagatcagaa gtactattcc agtatggacg attcaaggct tgcttcataa
      781 accaaggcaa gtaatagaga ttggagtctc taaaaaggta gttcctactg aatctaaggc
      841 catgcatgga gtctaagatt caaatcgagg atctaacaga actcgccgtg aagactggcg
      901 aacagttcat acagagtctt ttacgactca atgacaagaa gaaaatcttc gtcaacatgg
      961 tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagaccaaa
     1021 gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc
     1081 cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc
     1141 atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag
     1201 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     1261 agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc
     1321 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acacgcacga
     1381 gatctgcagg tcgactctag aggatcccct cgaggcgcgc caagctatca aacaagtttg
     1441 tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
     1501 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg
     1561 ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt
     1621 tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     1681 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     1741 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     1801 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     1861 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     1921 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     1981 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     2041 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     2101 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     2161 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     2221 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     2281 atgaattaca acagtactgc gatgagtggc aggcggggcg taatctagag gatccggctt
     2341 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     2401 tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca
     2461 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     2521 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     2581 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     2641 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     2701 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     2761 gtgatccccc tggccagtgc acgtctgctg tcagataaag tcccccgtga actttacccg
     2821 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     2881 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     2941 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct
     3001 gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     3061 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     3121 cttgtacaaa gtggttcgat aattccttaa ttaactagtt ctagagcggc cgcccaccgc
     3181 ggtggagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga
     3241 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     3301 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     3361 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     3421 tatcgcgcgc ggtgtcatct atgttactag atcgggaatt cgtaatcatg gtcatagctg
     3481 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3541 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3601 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5641 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5701 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5761 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5821 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     5881 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     5941 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6001 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6061 tcacgacgtt gtaaaacgac ggccagtgcc
//
