LOCUS       sequence-62389-e        5007 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 5007)
  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 5007)
  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: pZHY402
FEATURES             Location/Qualifiers
     source          1..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        488..833
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     776..796
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             854..874
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(926..947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1669..1920
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVMLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMEELYK"
     primer_bind     1857..1878
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1960..2212
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2233..2252)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2234..2252)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2262..2279)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2262..2278)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2271..2293)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2419..2874)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2556..2577)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2768..2787
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2901..3005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3006..3074,3075..3866)
                     /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     complement(3224..3243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4037..4625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4526..4545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4779..4796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4913..4934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4949..4954,4955..4972,4973..4979)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4987..5003
                     /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)."
ORIGIN
        1 agatttgcct tttcaatttc agaaagaatg ctaacccaca gatggttaga gaggcttacg
       61 cagcaggtct catcaagacg atctacccga gcaataatct ccaggaaatc aaataccttc
      121 ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga
      181 aagatatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc
      241 acaaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttccc actgaatcaa
      301 aggccatgga gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg
      361 aacagttcat acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg
      421 tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa
      481 gggcaattga gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc
      541 cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc
      601 atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag
      661 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
      721 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
      781 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagaga acacggggga
      841 ctctagaatg gctcctaaga agaagagaaa ggttataaca atggtgagca agggcgagga
      901 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      961 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1021 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     1081 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1141 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1201 caagacagat ctggcagttt catcatgaat gtgcaggagc tagcaactat taaggtggag
     1261 aatctcccac tagtcggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1321 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1381 accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     1441 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1501 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacaca
     1561 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1621 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1681 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1741 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgatgct gcccgacaac
     1801 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1861 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggaaga gctgtacaag
     1921 ccgcggttcc cgggagacct ttagagctcg aatttccccg atcgttcaaa catttggcaa
     1981 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     2041 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     2101 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     2161 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgtgggccc
     2221 agctccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     2281 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2341 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2401 agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2461 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2521 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2581 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2641 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2701 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2761 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2821 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2881 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2941 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3001 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3061 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3121 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3181 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3241 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3301 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3361 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3421 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3481 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3541 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3601 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3661 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3721 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3781 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3841 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3901 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3961 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4021 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4081 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4141 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4201 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4261 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4321 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4381 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4441 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4501 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4561 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4621 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4681 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4741 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4801 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4861 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     4921 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     4981 gtggaattgt gagcggataa caatttc
//
