LOCUS       sequence-11344-e        3998 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 3998)
  AUTHORS   Aoyama M, Agari K, Sun-Wada GH, Futai M, Wada Y
  TITLE     Simple and straightforward construction of a mouse gene targeting 
            vector using in vitro transposition reactions.
  JOURNAL   Nucleic Acids Res. 2005  . 33(5):e52.
  PUBMED    15784610
REFERENCE   2  (bases 1 to 3998)
  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: "Nucleic 
            Acids Res. 2005  . 33(5):e52."
COMMENT     Sequence Label: pBSDT-AII
FEATURES             Location/Qualifiers
     source          1..3998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          775..840
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(917..1489)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     primer_bind     complement(1810..1830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1810..1828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1849..1865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1849..1865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1862..1884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1873..1889
                     /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(1897..1903,1904..1921,1922..1927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1942..1963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2080..2097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2251..2839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2331..2350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3010..3801,3802..3870))
                     /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     3633..3652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3871..3975)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctgtt
      661 taaacggacc gcggtggcgg ccgctctagg atccggccgg cccgggcgat cccccgactc
      721 tagtggatct gcattccacc actgctccca ttcatcagtt ccataggttg gaatctaaaa
      781 tacacaaaca attagaatca gtagtttaac acattataca cttaaaaatt ttatatttac
      841 cttagagctt taaatctctg taggtagttt gtccaattat gtcacaccac agaagtaagg
      901 ttccttcaca aagagatcgc ctgacacgat ttcctgcaca ggcttgagcc atatactcat
      961 acatcgcatc ttggccacgt tttccacggg tttcaaaatt aatctcaagt tctacgctta
     1021 acgctttcgc ctgttcccag ttattaatat attcaacgct agaactcccc tcagcgaagg
     1081 gaaggctgag cactacacgc gaagcaccat caccgaacct tttgataaac tcttccgttc
     1141 cgacttgctc catcaacggt tcagtgagac ttaaacctaa ctctttctta atagtttcgg
     1201 cattatccac ttttagtgcg agaaccttcg tcagtcctgg atacgtcact ttgaccacgc
     1261 ctccagcttt tccagagagc gggttttcat tatctacaga gtatcccgca gcgtcgtatt
     1321 tattgtcggt actataaaac cctttccaat catcgtcata atttccttgt gtaccagatt
     1381 ttggcttttg tatacctttt tgaatggaat ctacataacc aggtttagtc ccgtggtacg
     1441 aagaaaagtt ttccatcaca aaagatttag aagaatcaac aacatcatca gggtccatgg
     1501 cgaggacgtg cagggtcgct cggtgttcga ggccacacgc gtcaccttaa tatgcgaagt
     1561 ggacctggga ccgcgccgcc ccgactgcat ctgcgtgttc gaattcgcca atgacaagac
     1621 gctgggcggg gtttgctcga cattgggtgg aaacattcca ggcctgggtg gagaggcttt
     1681 ttgcttcctc ttgcaaaacc acactgctcg acattgggtg gaaacattcc aggcctgggt
     1741 ggagaggctt tttgcttcct cttgaaaacc acactgctcg agggggggcc cggtacccag
     1801 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     2701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2821 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2881 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2941 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3001 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3061 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3961 aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
