LOCUS       sequence-36630-e        5122 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 5122)
  AUTHORS   Petersen LK, Stowers RS
  TITLE     A Gateway MultiSite Recombination Cloning Toolkit.
  JOURNAL   PLoS One. 2011;6(9):e24531. Epub 2011 Sep 9.
  PUBMED    21931740
REFERENCE   2  (bases 1 to 5122)
  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: "2011"; volume: "6(9)"; pages: "e24531. Epub 2011 Sep 9"
COMMENT     Sequence Label: pENTR R4-QF-R3
FEATURES             Location/Qualifiers
     source          1..5122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    3185..3308
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3363..3382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3364..3382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3387..3403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3387..3403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3505..3524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3516..4325
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3583..3602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4472..5060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4961..4980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttggcca acatgccgcc taaacgcaag acactcaatg ccgctgccga agccaatgcc
      781 cacgctgatg gccatgctga tggcaatgct gatggtcatg tcgctaacac tgcagcaagc
      841 agcaacaacg cccgttttgc ggacttgacc aacattgaca cacccggcct cggccctacc
      901 acgacgacgt tacttgtcga gcccgctcgt tcgaaacgcc agagagtctc gagggcctgt
      961 gatcagtgtc gagctgcacg tgaaaagtgt gatggaatcc agccggcttg cttcccctgt
     1021 gtgtcgcagg gccggtcgtg tacctaccag gccagtccca agaagcgagg agtccagacg
     1081 ggctacatcc gcactctcga actggctctg gcttggatgt tcgagaacgt tgcccgcagc
     1141 gaggacgccc tccacaatct tttggtccgt gatgctggcc agggcagcgc tctcctggtc
     1201 ggcaaagact cgcctgctgc agaacgcctg catgcaagat gggcgacgag tcgagtcaac
     1261 aaaagcatca cccgtcttct ctcaggtcag gccgcacaag atccatctga agacggccaa
     1321 tccccgtccg aagacataaa tgtccaagat gccggggcaa agacatccga cttccctcat
     1381 gcgcctcact tgactttctc ggcgcctaag tctagtacag ctgagacacg cactctacca
     1441 ggcccggtcc gaccacctat ttcggcaaac accctggaaa acaaccttca gccagatggt
     1501 accgggatag ggaagctacc acccaatcat tggcgcctgc tggatatcta cttttcctac
     1561 acgcattctt ggctccctat cctcgagaag aaagacatgt accaagcatt gtaccagtac
     1621 tctgaacaag gctcgttgct tccctctgcg aatgtcgagt ctggcgttca tgccgagctc
     1681 tggagcgcgc tcgccctggc gtccttccag gctgctgcta ctgctgcatc gagtgctacg
     1741 ggtccagctt cagctgctca tggccatgac aatgccatca atccttcacc tgcagacata
     1801 tctgacacag cccgaaagct catacctttg gaaagcgggc cgttccaggt tcagcactgc
     1861 agagcgttgc tgcttctttg tctcgtaagc cttgggcggg atgattggga gtctgcttgg
     1921 ttgctggttg gctttgcggt ccgcgtccta cttgttgttc gcacccagtt gcctcctgat
     1981 gatgaccggc cacgaccaag aatgcgtgcg ctgctcgtcg cgtgcttcat cgtggatacc
     2041 attgtgtcta tgagacacaa cgtgccggcc catctcaagc cagacgacat tgcggatctg
     2101 ccgttacctg aagacggtca agatcaatgg gagccgtgga caccatgtga gggcttaggc
     2161 ggtgaacaca ccatgctgca aatgttgagg aacccggcat accctttaag cacattcaac
     2221 cacctatatg gcgtgaccaa gctggttgct ttggagcttc tgccaagaat acgaacatct
     2281 tcacagaacg ctcccttgga gttcaggtcg cggttgcagc aggtaatcgg ccacaattct
     2341 cccttcagcg tctttgtcct ttcccaggat acagcatcgg cttttgtgcc tactgcatac
     2401 cttacccgta ccgtttattt atgggcagct gccttttctg agcctctcaa cgaacactac
     2461 tcgcatcttc tgatcgaaac tcttgatcag tatcagaagc ggtttggtac atatgcaatc
     2521 ccacctctga tcccttctct tctagactcc cttcttgctt taaagaaaca atcacattct
     2581 tcagagcggc atcgaaggca cttggaagag cttttccccg cctactcctc catttggcct
     2641 cggggaggcc gacacagcaa tactggcctc caacccatac gacaacttga gcttccgccg
     2701 actgcgactg ccactgcaag tatcatgccc catgtcatgg aacagcccct gtcaacgtca
     2761 ataaatccgg tcaatgatcg gtttaatgga ataccgaatc ctacccccta caatagcgat
     2821 gcggccctag acgcaattac tcaaaccaat gactacggat cagtcaatac ccatggcatt
     2881 ctcagtacat atccaccacc tgccacccac ttgaacgagg cctcggtggc ccttgcgcct
     2941 ggaggtgctc ctcccagacc acccccgcca tacgttgata gtacaacgaa ccatcctcct
     3001 taccatagta acctcgtccc catggccaat ttcggatatt cgactgtaga ctatgatgcg
     3061 atggtggatg atctggcatc gatcgagtac acggatgcgg ttgacgttga tccgcagttc
     3121 atgacgaacc tcgggtttgt cccagggtgt aattttagcg atatcaacac gtatgagcaa
     3181 tagacaactt tgtataataa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     3241 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     3301 tcactatgaa tcaactactt agatggtatt agtgacctgt actgcagggc ggccgcgata
     3361 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     3421 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3481 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3541 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3601 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     3661 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3721 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3781 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     3841 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3901 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     3961 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     4021 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     4081 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     4141 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     4201 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     4261 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     4321 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     4381 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     4441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5101 gctggccttt tgctcacatg tt
//
