LOCUS       sequence-84676-e        4614 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 4614)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 4614)
  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: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LI F-G Hygro c1
FEATURES             Location/Qualifiers
     source          1..4614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..48)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(32..48)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(45..67)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    56..72
                     /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(80..86,87..104,105..110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    125..146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(263..280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(434..1022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(514..533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1137..1142,1143..1159,1160..1165)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1354..1887
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2379..2396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2380..2396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2965..3310
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3253..3273
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             3317..4342
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    4383..4557
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
       61 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      121 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      181 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      241 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      301 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      361 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      421 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      481 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      541 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      601 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      661 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      721 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      781 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      841 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      901 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      961 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1021 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1081 aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga
     1141 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt
     1201 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     1261 atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     1321 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg
     1381 cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca
     1441 tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt
     1501 gagttcggcg acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac
     1561 ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc
     1621 gctctcgcgg cttacgttct gcccaggttt gagcagccgc gtagtgagat ctatatctat
     1681 gatctcgcag tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc
     1741 ctcaagcatg aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt
     1801 gacgatcccg cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt
     1861 gatatcgacc caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc
     1921 gcggagttgt tcggtaaatt gtcacaacgc cgctcatgac attaacctat aaaaataggc
     1981 gtatcacgag gcccttttgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagaaggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2401 tcgagctcgg tacctcgcga atgcatctag atatcggatc ccgggcccgt cgactgcaga
     2461 ggtacgggtc tcatgagaga ttagcctttt caatttcaga aagaatgcta acccacagat
     2521 ggttagagag gcttacgcag cacgactcat caagacgatc tacccgagca ataatctcca
     2581 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
     2641 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
     2701 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
     2761 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
     2821 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
     2881 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
     2941 agtctcagaa gagcaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
     3001 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
     3061 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
     3121 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaacacgt
     3181 tccaaccacg acttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     3241 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     3301 ggagagaaca cgcaccatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     3361 gatcgaaaag ttcgacagcg tgtccgacct gatgcagctc tcggagggcg aagaatctcg
     3421 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3481 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     3541 ggaagtgctt gacattgggg agtttagcga gagcctgacc tattgcatct cccgccgttc
     3601 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tacaaccggt
     3661 cgcggaggct atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     3721 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     3781 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     3841 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     3901 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     3961 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4021 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4081 gcttgcagga tcgccacgac tccgggcgta tatgctccgc attggtcttg accaactcta
     4141 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4201 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     4261 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     4321 tcgtccgagg gcaaagaaat agagtagatg ccgaccggga tctgtcgatc gacaagctcg
     4381 agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg
     4441 tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac
     4501 ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcccc
     4561 cgaattaatt cggcgttaat tcagtacatt aaaaacgtcc gcatgtctga gacc
//
