LOCUS       sequence-436166-        3137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing ITR sequences compatible with Minos transposase 
            and multiple cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3137)
  TITLE     Stable germline transgenesis using the Minos Tc1/mariner element in 
            the sea urchin, Lytechinus pictus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3137)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Minos ITR loxp MCS lox2272
FEATURES             Location/Qualifiers
     source          1..3137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(340..373)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(540..573)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(973..989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(973..989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(986..1008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1161..1749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1241..1260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1866..1890)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1990..2781,2782..2850))
                     /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     2613..2632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2851..2955)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3054..3076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3068..3085
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3069..3085
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcttctataa attatgccaa gcacgttagc agcttctacg agccccaacc actattaatt
       61 cgaacagcat gtttttttgc agtgcgcaat gtttaacaca ctatattatc aatactacta
      121 aagataacac ataccaatgc atttcgtctc aaagagaatt ttattctctt cacgacgaaa
      181 aaaaaagttt tgctctattt ccaacaacaa caaaaatatg agtaatttat tcaaacggtt
      241 tgcttaagag ataagaaaaa agtgaccact attaattcga acgcggcgta agctacctta
      301 atctcaagaa gagcaaaaca aaagcaacta atgtaacgga taacttcgta tagcatacat
      361 tatacgaagt tataaccata tgtgacgtct acggatccgc atgcactagt ggcgcgccat
      421 taaccatggt taaagcttaa ggccggccag cgatcgccgg acatcgattg gaattcaagg
      481 cctctcgagc ctctagaacg ttaacaaggt accggcggcc gccttcacgc gtggttacca
      541 taacttcgta taggatactt tatacgaagt tatccgcggg ggaaattaat taaattattg
      601 ttttaagtat gatagtaaat cacattacgc cgcgttcgaa ttaatagtgg tcactttttt
      661 cttatctctt aagcaaaccg tttgaataaa ttactcatat ttttgttgtt gttggaaata
      721 gagcaaaact ttttttttcg tcgtgaagag aataaaattc tctttgagac gaaatgcatt
      781 ggtatgtgtt atctttagta gtattgataa tatagtgtgt taaacattgc gcactgcaaa
      841 aaaaacatgc tgttcgaatt aatagtggtt ggggctcgta gaaaacgaaa aatatcttaa
      901 gctagcatag agaatgaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat
      961 gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa
     1021 tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg
     1081 gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc
     1801 gccgtcccgt caagtcagcg taaatgggta gggggcttca aatcgtcctc gtgataccaa
     1861 ttcggagcct gcttttttgt acaaacttgt tgataatggc aattcaagga tcttcaccta
     1921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2041 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2101 atctggcccc agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc
     2161 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2221 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2281 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2341 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2401 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2461 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2521 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2581 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2641 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2701 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2761 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2821 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2881 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2941 aataggggtt ccgcgcacat ttccccgaaa agtgccagat acctgaaaca aaacccatcg
     3001 tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt tttcccagtc
     3061 acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga ataaggaagt
     3121 gccattccgc ctgacct
//
