LOCUS       sequence-483538-        7403 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  For Minos transgenesis in insect, expresses the 3xP3::mCherry 
            eye/glia transgenesis, and the mBaoJin in Posterior Silk Glands 
            (under the control of the Fibroin-L promoter from Plodia 
            interpunctella).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  TITLE     Minos-mediated transgenesis in the pantry moth Plodia interpunctella
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pMi[3xP3::mCherry-SV40, Plodia-FibL::mBaojin-P10]
FEATURES             Location/Qualifiers
     source          1..7403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..35)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(17..35)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(42..59)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(42..58)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(51..73)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      200..655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(287..306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     497..518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        681..785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(786..854,855..1646)
                     /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(1004..1023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1817..2405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2306..2325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2559..2576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2693..2714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2729..2734,2735..2752,2753..2759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2767..2783
                     /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     2772..2794
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2791..2807
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2791..2807
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2828..2846
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3329..3345
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3330..3346
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3631..3640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3637..4347
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3782..3800)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4050..4069
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4474..4595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4511..4530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4565..4584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             5615..6316
                     /codon_start=1
                     /product="bright and photostable monomeric green 
                     fluorescent protein derived from StayGold (Zhang et al., 
                     2024)"
                     /label=mBaoJin
                     /label=mStayGold
                     /note="/vntifkey=21"
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYTQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     primer_bind     complement(7001..7017)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7001..7017)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gcgagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
       61 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      121 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      181 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      241 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
      661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2581 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2641 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2701 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2761 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2821 agctcgaaat taaccctcac taaagggaac aaaagctggt accaagtgct tgaaatgcta
     2881 aatgttttca atttttcgcc attaagacaa gcctacacaa atgcttctat aaattatgcc
     2941 aagcacgtta gcagcttcta cgagccccaa ccactattaa ttcgaacagc atgttttttt
     3001 gcagtgcgca atgtttaaca cactatatta tcaatactac taaagataac acataccaat
     3061 gcatttcgtc tcaaagagaa ttttattctc ttcacgacga aaaaaaaagt tttgctctat
     3121 ttccaacaac aacaaaaata tgagtaattt attcaaacgg tttgcttaag agataagaaa
     3181 aaagtgacca ctattaattc gaacgcggcg taagcttacc ttaatctcaa gaagagcaaa
     3241 acaaaagcaa ctaatgtaac ggaatcatta tctagttatg atctgcaaat aatgctgcag
     3301 agctggggga tccccggcgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc
     3361 gaattcgagc tcgcccgggg atctaattca attagagact aattcaatta gagctaattc
     3421 aattaggatc caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc
     3481 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
     3541 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggggtacc gctagagtcg
     3601 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat
     3661 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3721 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3781 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3841 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3901 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3961 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     4021 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     4081 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     4141 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     4201 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     4261 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     4321 ggcggcatgg acgagctgta caagtaaagc ggccggccgc gactctagat cataatcagc
     4381 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4441 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4501 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4561 agttgtggtt tgtccaaact catcaatgta tcttagctag ctattgaatg caataagatg
     4621 tcacaataca ataaaagctt gtgtcaaaaa tggtatattt attaaaaaaa ttattctctt
     4681 aagactgcta tccatcatct aaatatacct ataaattaat ccgagtaaaa catctatact
     4741 tatctaattt ttattgaaaa tctttacgat acaatagaag actgaatttg actttgtctt
     4801 atccggcttt gttttgtaat ttagagattt tatgcgattt cagcgaagat acgggtgtaa
     4861 ttcgcaacaa agatggtaat atatcgtaaa aatgcttagc cttttatgat tgagttgacg
     4921 gtgctcgttc gccgactttg tatgtttggg gaaaagataa aggtaaacaa ttaccagtgt
     4981 ccacaagtga ttgacaagtc tctcatcctc gcgtgttctc gattaaattt ggtgctggac
     5041 gccccgaagt ccaaagatcg tgtaaaacaa ggaacattta aattttgaag gtttggccgt
     5101 gctgatgttt agttagtccg agttgctctt ctaacctatt gacgtgtcta attgacgtgt
     5161 tagcaaatcc ttcattttta accttcaact ttatttactt aacaaatgac gttgcaagtg
     5221 aaaaatatct taatttcaca ttgtaccgaa aacaatttcc attaatgctc gcctttggac
     5281 aaacatataa acaaacgatt caattaatcg ctttgtacaa aatctaaact aaaacaatta
     5341 tatacctgca tttatataac aatcatcaat tcactttaaa ccttaatgca acagtaataa
     5401 gattcgcact taaaaattta tgaaataatg tcgtttcgtt ttcttgttaa tctggttatt
     5461 gttcataggg tgttatttga ccaggtgcaa attgtttgaa gattatttat atatattgtt
     5521 atattagtgt ataaatagta gataaagaca gacgattttg tgtataatat ataccgaatt
     5581 ggtcgaatta agaccagcaa atagtagtat caaaatggtc tcaaagggag aggaggaaaa
     5641 catggctagt acaccattta aatttcaact taaaggaacc atcaatggca aatcgtttac
     5701 cgttgaaggc gaaggtgaag ggaactcaca tgaaggttct cataaaggaa aatatgtttg
     5761 tacaagtgga aaactaccga tgtcatgggc agcacttggg acaacctttg gttatggaat
     5821 gaaatattat accaaatatc ctagtggact gaagaactgg tttcgtgaag taatgcccgg
     5881 aggctttacc tacgatcgtc atattcaata taaaggcgat gggagtatcc atgcaaaaca
     5941 ccaacacttt atgaaaaatg ggacttatca caacattgta gaatttaccg gtcaggattt
     6001 taaagaaaat agtccagtct taactggaga tatgaatgtc tcattaccga atgaagtccc
     6061 acaaataccc agagatgatg gagtagaatg cccagtgacc ttgctttatc ctttattatc
     6121 ggataaatca aaatacgtcg aggctcacca atatacaatc tgcaagcctc ttcataatca
     6181 accagcacct gatgtcccat atcactggat tcgtaaacaa tacacacaaa gcaaagatga
     6241 tgccgaggaa cgcgatcata tctgtcaatc agagactctc gaagcacact taaagggcat
     6301 ggacgagctg tataagtagc acgtgatgaa tcgtttttaa aataacaaat caattgtttt
     6361 ataatattcg tacgattctt tgattatgta ataaaatgtg atcattagga agattacgaa
     6421 aaatataaaa aatatgagtt ctgtgtgtat aacaaatgct gtaaacgcca caattgtgtt
     6481 tgttgcaaat aaacccatga ttatttgatt aaaattgttg ttttctttgt tcatagacaa
     6541 tagtgtgttt tgcctaaacg tgtactgcat aaactccatg cgagtgtata gcgagctagt
     6601 ggctaacgct tgccccacca aagtagattc gtcaaaatcc tcaatttcat caccctcctc
     6661 caagtttaac atttggccgt cggaattaac ttctaaagat gccacataat ctaataaatg
     6721 aaatagagat tcaaacgtgg cgtcatcgtc cgtttcgacc atttccgaaa agaactcggg
     6781 cataaactct atgatttctc tggacgtggt gttgtcgaaa ctctcaaagt acgcagtcag
     6841 gaacgtgcgc gacatgtcgt cgggaaactc gcgcggaaac atgttgttgt aaccgaacgg
     6901 gtcccatagc gccaaaacca aatctgccag cgtcaataga atgagcacga tgccgacaat
     6961 ggagctggct tggatagcga ttcgagttaa caacggccgg gatccactag ttctagagcg
     7021 gccgccaccg cgggacgtta cacaattcta atattaatta aattattgtt ttaagtatga
     7081 tagtaaatca cattacgccg cgttcgaatt aatagtggtc acttttttct tatctcttaa
     7141 gcaaaccgtt tgaataaatt actcatattt ttgttgttgt tggaaataga gcaaaacttt
     7201 ttttttcgtc gtgaagagaa taaaattctc tttgagacga aatgcattgg tatgtgttat
     7261 ctttagtagt attgataata tagtgtgtta aacattgcgc actgcaaaaa aaacatgctg
     7321 ttcgaattaa tagtggttgg ggctcgtaga aaacgaaaaa tatcttaagc tagcatagag
     7381 aatggagcaa aactcaattt gat
//
