LOCUS       sequence-84625-e        4239 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 4239)
  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 4239)
  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 neo c2
FEATURES             Location/Qualifiers
     source          1..4239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(24..29,30..46,47..52)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             241..774
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     861..879
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(917..939)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1039..1058
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1252..1274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1266..1283
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1267..1283
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1424..1603
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        1456..1639
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             1660..2454
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1714..1733)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2324..2343
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2844..3096
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3158..3174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3158..3174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3171..3193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3182..3198
                     /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(3206..3212,3213..3230,3231..3236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3251..3272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3389..3406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3560..4148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3640..3659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggatgaa ggcacgaacc cagttgacat aagcctgttc ggttcgtaaa ctgtaatgca
       61 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
      121 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct atgcctcggg
      181 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      241 atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaggt
      301 ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg
      361 cgggctgctc ttgatctttt cggtcgtgag ttcggcgacg tagccaccta ctcccaacat
      421 cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct
      481 gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caggtttgag
      541 cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca ccggaggcag
      601 ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat
      661 gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg
      721 ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaacaattc
      781 gttcaagccg agatcggctt cccggccgcg gagttgttcg gtaaattgtc acaacgccgc
      841 tcatgacatt aacctataaa aataggcgta tcacgaggcc cttttgtctc gcgcgtttcg
      901 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gaaggtcaca gcttgtctgt
      961 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1021 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     1081 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     1141 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1201 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1261 gacgttgtaa aacgacggcc agtgaattcg agctcggtac ctcgcgaatg catctagata
     1321 tcggatcccg gtctcttgag gatcatgagc ggagaattaa gggagtcacg ttatgacccc
     1381 cgccgatgac gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag
     1441 gagccactca gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat
     1501 tattgcgcgt tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa
     1561 atgctccact gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc
     1621 tcaatccaaa taatctgcac cggatctgga tcgtttcgca tgattgaaca agatggattg
     1681 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1741 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1801 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     1861 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     1921 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     1981 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2041 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2101 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     2161 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc
     2221 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     2281 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     2341 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     2401 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     2461 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     2521 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     2581 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccacgggatc tctgcggaac
     2641 aggcggtcga aggtgccgat atcattacga cagcaacggc cgacaagcac aacgccacga
     2701 tcctgagcga caatatgatc gggcccggcg tccacatcaa cggcgtcggc ggcgactgcc
     2761 caggcaagac cgagatgcac cgcgatatct tgctgcgttc ggatattttc gtggagttcc
     2821 cgccacagac ccggatgatc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
     2881 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     2941 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     3001 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     3061 aattatcgcg cgcggtgtca tctatgttac tagatcgggt gtctgagacc gggcccgtcg
     3121 actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3181 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3241 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3301 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3361 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3421 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3481 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3541 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3601 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3661 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3721 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3781 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3841 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3901 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3961 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4021 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4081 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4141 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4201 cacgttaagg gattttggtc atgaggcgca caccgtgga
//
