LOCUS       sequence-207451-        5043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transgenesis vector based on the Minos transposon, carrying the 
            3xP3-EGFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5043)
  AUTHORS   Pavlopoulos A, Berghammer AJ, Averof M, Klingler M
  TITLE     Efficient transformation of the beetle Tribolium castaneum using the
            Minos transposable element: quantitative and qualitative analysis of
            genomic integration events.
  JOURNAL   Genetics. 2004 Jun;167(2):737-46.
  PUBMED    15238525
REFERENCE   2  (bases 1 to 5043)
  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: "Genetics.";
            date: "2004-06"; volume: "167(2)"; pages: "737-46"
COMMENT     Sequence Label: pMi(3xP3-EGFP)
FEATURES             Location/Qualifiers
     source          1..5043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..487
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     472..488
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      773..782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(779..781,782..784,785..1498)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(779..781,782..784,785..1495)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(824..845)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1085..1104)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1432..1453
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1625..1746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1662..1681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1716..1735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1783..1799)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1783..1799)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2201..2220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2202..2220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2227..2244)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2227..2243)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2236..2258)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2385..2840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2472..2491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2682..2703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2866..2970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2971..3039,3040..3831)
                     /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     complement(3189..3208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4002..4590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4491..4510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4744..4761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4878..4899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4914..4919,4920..4937,4938..4944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4952..4968
                     /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)."
     primer_bind     4957..4979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4976..4992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4976..4992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5013..5031
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gtaccaagtg cttgaaatgc taaatgtttt caatttttcg ccattaagac aagcctacac
       61 aaatgcttct ataaattatg ccaagcacgt tagcagcttc tacgagcccc aaccactatt
      121 aattcgaaca gcatgttttt ttgcagtgcg caatgtttaa cacactatat tatcaatact
      181 actaaagata acacatacca atgcatttcg tctcaaagag aattttattc tcttcacgac
      241 gaaaaaaaaa gttttgctct atttccaaca acaacaaaaa tatgagtaat ttattcaaac
      301 ggtttgctta agagataaga aaaaagtgac cactattaat tcgaacgcgg cgtaagctta
      361 ccttaatctc aagaagagca aaacaaaagc aactaatgta acggaatcat tatctagtta
      421 tgatctgcaa ataatgctgc agagctgggg gatccccggc gggccccccc tcgaggtcga
      481 cggtatcgat aagcttgata tcgaattcga gctcgcccgg ggatctaatt caattagaga
      541 ctaattcaat tagagctaat tcaattagga tccaagctta tcgatttcga accctcgacc
      601 gccggagtat aaatagaggc gcttcgtcta cggagcgaca attcaattca aacaagcaaa
      661 gtgaacacgt cgctaagcga aagctaagca aataaacaag cgcagctgaa caagctaaac
      721 aatcggggta ccgctagagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
      781 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      841 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      901 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      961 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1021 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1081 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1141 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1201 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1261 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1321 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1381 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1441 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     1501 cggccggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1561 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1621 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1741 atcttaaagc ttatcgatac gcgtacggcg cgcctagggg gggatccact agttctagag
     1801 cggccgccac cgcgggacgt tacacaattc taatattaat taaattattg ttttaagtat
     1861 gatagtaaat cacattacgc cgcgttcgaa ttaatagtgg tcactttttt cttatctctt
     1921 aagcaaaccg tttgaataaa ttactcatat ttttgttgtt gttggaaata gagcaaaact
     1981 ttttttttcg tcgtgaagag aataaaattc tctttgagac gaaatgcatt ggtatgtgtt
     2041 atctttagta gtattgataa tatagtgtgt taaacattgc gcactgcaaa aaaaacatgc
     2101 tgttcgaatt aatagtggtt ggggctcgta gaaaacgaaa aatatcttaa gctagcatag
     2161 agaatggagc aaaactcaat ttgatgcgag ctccaattcg ccctatagtg agtcgtatta
     2221 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2281 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2341 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2461 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2581 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2701 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2761 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2821 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2941 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3001 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3061 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3121 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3181 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3241 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3301 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3361 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3421 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3481 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3541 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3601 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3661 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3721 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3781 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3841 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3901 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4621 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4681 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4741 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4801 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4861 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     4921 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4981 aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag
     5041 ctg
//
