LOCUS       sequence-36626-e        5012 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 5012)
  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 5012)
  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 L5-QF-L2
FEATURES             Location/Qualifiers
     source          1..5012
                     /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    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3124..3223)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3253..3272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3254..3272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3277..3293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3277..3293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3395..3414)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3406..4215
                     /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(3473..3492)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4362..4950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4851..4870
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttggccaa catgccgcct aaacgcaaga cactcaatgc cgctgccgaa gccaatgccc
      721 acgctgatgg ccatgctgat ggcaatgctg atggtcatgt cgctaacact gcagcaagca
      781 gcaacaacgc ccgttttgcg gacttgacca acattgacac acccggcctc ggccctacca
      841 cgacgacgtt acttgtcgag cccgctcgtt cgaaacgcca gagagtctcg agggcctgtg
      901 atcagtgtcg agctgcacgt gaaaagtgtg atggaatcca gccggcttgc ttcccctgtg
      961 tgtcgcaggg ccggtcgtgt acctaccagg ccagtcccaa gaagcgagga gtccagacgg
     1021 gctacatccg cactctcgaa ctggctctgg cttggatgtt cgagaacgtt gcccgcagcg
     1081 aggacgccct ccacaatctt ttggtccgtg atgctggcca gggcagcgct ctcctggtcg
     1141 gcaaagactc gcctgctgca gaacgcctgc atgcaagatg ggcgacgagt cgagtcaaca
     1201 aaagcatcac ccgtcttctc tcaggtcagg ccgcacaaga tccatctgaa gacggccaat
     1261 ccccgtccga agacataaat gtccaagatg ccggggcaaa gacatccgac ttccctcatg
     1321 cgcctcactt gactttctcg gcgcctaagt ctagtacagc tgagacacgc actctaccag
     1381 gcccggtccg accacctatt tcggcaaaca ccctggaaaa caaccttcag ccagatggta
     1441 ccgggatagg gaagctacca cccaatcatt ggcgcctgct ggatatctac ttttcctaca
     1501 cgcattcttg gctccctatc ctcgagaaga aagacatgta ccaagcattg taccagtact
     1561 ctgaacaagg ctcgttgctt ccctctgcga atgtcgagtc tggcgttcat gccgagctct
     1621 ggagcgcgct cgccctggcg tccttccagg ctgctgctac tgctgcatcg agtgctacgg
     1681 gtccagcttc agctgctcat ggccatgaca atgccatcaa tccttcacct gcagacatat
     1741 ctgacacagc ccgaaagctc atacctttgg aaagcgggcc gttccaggtt cagcactgca
     1801 gagcgttgct gcttctttgt ctcgtaagcc ttgggcggga tgattgggag tctgcttggt
     1861 tgctggttgg ctttgcggtc cgcgtcctac ttgttgttcg cacccagttg cctcctgatg
     1921 atgaccggcc acgaccaaga atgcgtgcgc tgctcgtcgc gtgcttcatc gtggatacca
     1981 ttgtgtctat gagacacaac gtgccggccc atctcaagcc agacgacatt gcggatctgc
     2041 cgttacctga agacggtcaa gatcaatggg agccgtggac accatgtgag ggcttaggcg
     2101 gtgaacacac catgctgcaa atgttgagga acccggcata ccctttaagc acattcaacc
     2161 acctatatgg cgtgaccaag ctggttgctt tggagcttct gccaagaata cgaacatctt
     2221 cacagaacgc tcccttggag ttcaggtcgc ggttgcagca ggtaatcggc cacaattctc
     2281 ccttcagcgt ctttgtcctt tcccaggata cagcatcggc ttttgtgcct actgcatacc
     2341 ttacccgtac cgtttattta tgggcagctg ccttttctga gcctctcaac gaacactact
     2401 cgcatcttct gatcgaaact cttgatcagt atcagaagcg gtttggtaca tatgcaatcc
     2461 cacctctgat cccttctctt ctagactccc ttcttgcttt aaagaaacaa tcacattctt
     2521 cagagcggca tcgaaggcac ttggaagagc ttttccccgc ctactcctcc atttggcctc
     2581 ggggaggccg acacagcaat actggcctcc aacccatacg acaacttgag cttccgccga
     2641 ctgcgactgc cactgcaagt atcatgcccc atgtcatgga acagcccctg tcaacgtcaa
     2701 taaatccggt caatgatcgg tttaatggaa taccgaatcc taccccctac aatagcgatg
     2761 cggccctaga cgcaattact caaaccaatg actacggatc agtcaatacc catggcattc
     2821 tcagtacata tccaccacct gccacccact tgaacgaggc ctcggtggcc cttgcgcctg
     2881 gaggtgctcc tcccagacca cccccgccat acgttgatag tacaacgaac catcctcctt
     2941 accatagtaa cctcgtcccc atggccaatt tcggatattc gactgtagac tatgatgcga
     3001 tggtggatga tctggcatcg atcgagtaca cggatgcggt tgacgttgat ccgcagttca
     3061 tgacgaacct cgggtttgtc ccagggtgta attttagcga tatcaacacg tatgagcaat
     3121 agtacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     3181 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc
     3241 ggccgcgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     3301 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     3361 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     3421 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     3481 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     3541 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     3601 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     3661 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     3721 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     3781 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     3841 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     3901 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     3961 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     4021 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     4081 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     4141 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     4201 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     4261 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     4321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4501 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4981 ctggcctttt gctggccttt tgctcacatg tt
//
