LOCUS       sequence-202023-        3459 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 3459)
  AUTHORS   Balciunas D, Wangensteen KJ, Wilber A, Bell J, Geurts A, Sivasubbu 
            S, Wang X, Hackett PB, Largaespada DA, McIvor RS, Ekker SC
  TITLE     Harnessing a high cargo-capacity transposon for genetic applications
            in vertebrates.
  JOURNAL   PLoS Genet. 2006 Nov 10;2(11):e169. Epub 2006 Aug 28.
  PUBMED    17096595
REFERENCE   2  (bases 1 to 3459)
  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 
            Genet."; date: "2006-11-10"; volume: "2(11)"; pages: "e169. Epub 
            2006 Aug 28"
COMMENT     Sequence Label: pminiTol2
FEATURES             Location/Qualifiers
     source          1..3459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(2..13,14..201)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     misc_feature    join(346..508,509..520)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        complement(571..589)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(572..589)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(607..623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(620..642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    631..647
                     /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(655..661,662..679,680..685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    700..721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(838..855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1009..1597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1089..1108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1768..2559,2560..2628))
                     /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     2391..2410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2629..2733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2811..3266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2948..2969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3160..3179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3392..3414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3406..3423
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3407..3423
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3430..3449
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3430..3448
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccagaggtgt aaagtacttg agtaatttta cttgattact gtacttaagt attatttttg
       61 gggattttta ctttacttga gtacaattaa aaatcaatac ttttactttt acttaattac
      121 atttttttag aaaaaaaagt actttttact ccttacaatt ttatttacag tcaaaaagta
      181 cttatttttt ggagatcact tcattctatt ttcccttgct attaccaaac caattgaatt
      241 gcgctgatgc ccagtttaat ttaaatagat ctggccatct agagcggccg cgcgcactag
      301 tgaattccat ggatatcaag cttaaacaag aatctctagt tttctttctt gcttttactt
      361 ttacttcctt aatactcaag tacaatttta atggagtact tttttacttt tactcaagta
      421 agattctagc cagatacttt tacttttaat tgagtaaaat tttccctaag tacttgtact
      481 ttcacttgag taaaattttt gagtactttt tacacctctg ctcgaccata tgggagagct
      541 cccaacgcgt tggatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta
      601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
      841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
      901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2761 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     2821 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2881 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2941 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3001 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3061 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3121 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3181 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3241 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     3301 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     3361 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3421 agtgaattgt aatacgactc actatagggc gaattgggc
//
