LOCUS       Exported                7634 bp ds-DNA     circular SYN 20-NOV-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7634)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Nov 20, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7634
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    152..771
                     /label=pBBR322 Origin
                     /note="color: #75c6a9"
     primer_bind     1055..1074
                     /label=BC108
                     /note="color: black; sequence: agctcactcattaggcaccc"
     misc_feature    complement(1144..1170)
                     /label=mzw123
                     /note="color: #f58a5e; direction: LEFT"
     misc_feature    1171..1188
                     /label=mzw188
                     /note="color: #b7e6d7"
     misc_feature    1189..1511
                     /label=5'piggyBac*
                     /note="color: #85dae9"
     primer_bind     1251..1275
                     /label=BC98
                     /note="color: black; sequence: tttctaaatagcgcgaatccgtcgc"
     misc_feature    complement(1512..1552)
                     /label=mzw119
                     /note="color: #ffef86; direction: LEFT"
     misc_feature    1533..1589
                     /label=mzw129
                     /note="color: #b4abac"
     misc_feature    1553..1589
                     /label=mzw116
                     /note="color: #c7b0e3"
     primer_bind     1565..1609
                     /label=BC78
                     /note="color: black; sequence: 
                     cttgtactgtcgaccctgtggaatgaccgtcgacctcgacggatc"
     primer_bind     complement(1570..1594)
                     /label=BC79
                     /note="color: black; sequence: acggtcattccacagggtcgacagt"
     promoter        1590..2965
                     /label=hEF1a Promoter
                     /note="color: #d59687"
     primer_bind     2619..2638
                     /label=BC59
                     /note="color: black; sequence: agccgtcgcttcatgtgact"
     misc_feature    2970..2994
                     /label=B1
                     /note="color: #d6b295"
     primer_bind     complement(2981..3003)
                     /label=BC56
                     /note="color: black; sequence: ggtcatagcagcctgcttttttg"
     misc_feature    3001..3003
                     /label=Min Koz
                     /note="color: #f58a5e"
     CDS             3004..4521
                     /label=CRY2
                     /note="color: #b4abac; direction: RIGHT"
     primer_bind     complement(4499..4521)
                     /label=BC65
                     /note="color: black; sequence: 
                     AAAGCTCCTCCCCCTTGCTGACCATggtggcgaccggtggatcccggg"
     primer_bind     complement(4507..4521)
                     /label=RL 144
                     /note="color: black; sequence: 
                     AAAGCTCCTCCCCCTTGCTGACCATggtggcgaccggtgg"
     misc_feature    4516..4521
                     /label=miscellaneous
                     /note="color: #b4abac"
     CDS             4522..5208
                     /label=LRP6
                     /note="color: #b4abac; direction: RIGHT"
     primer_bind     4522..4544
                     /label=RL 147
                     /note="color: black; sequence: 
                     CCTCGGAATGGATGAGCTCTATAAGgatggtggaggaggtagtggagg"
     primer_bind     complement(5186..5260)
                     /label=HL 32
                     /note="color: black; sequence: 
                     TACCCTTTGAACTGCCTGAGCCACTCCCGGAGCTTCCTCCGGATCCAGAACTggaggag
                     tctgtacagggagagg; added: 2021-08-12"
     primer_bind     5236..5325
                     /label=HL 31
                     /note="color: black; sequence: 
                     AGTGGCTCAGGCAGTTCAAAGGGTAAAAAGAAGAAAAAGAAGTCAAAGACAAAGTGTGT
                     AATTATGggaagcggagctactaacttcagc; added: 2021-08-12"
     misc_feature    5302..5367
                     /label=2A
                     /note="color: #d6b295"
     CDS             5302..5367
                     /label=Translation 5209-5274
                     /note="color: #993366; direction: RIGHT"
     primer_bind     5322..5387
                     /label=BC80
                     /note="color: black; sequence: 
                     cagcctgctgaagcaggctggagacgtggaggagaaccctggacctatgaccgagtaca
                     agcccac"
     misc_feature    5368..5986
                     /label=Puromycin
                     /note="color: #f58a5e"
     primer_bind     5368..5388
                     /label=BC5 -overlap
                     /note="color: black; sequence: atgaccgagtacaagcccacg"
     CDS             5703..5705
                     /label=Translation 5610-5612
                     /note="color: #993366; direction: RIGHT"
     misc_feature    5987..6004
                     /label=start
                     /note="color: #b7e6d7"
     primer_bind     complement(6011..6030)
                     /label=BC100
                     /note="color: black; sequence: cgtgcgctcctttcggtcgg"
     misc_feature    6053..6300
                     /label=3'_PibbgyBack
                     /note="color: #c6c9d1"
     misc_feature    complement(6305..6348)
                     /label=mzw117
                     /note="color: #9eafd2; direction: LEFT"
     misc_feature    6327..6351
                     /label=mzw122
                     /note="color: #84b0dc"
     primer_bind     complement(6391..6410)
                     /label=BC109
                     /note="color: black; sequence: gcagattgtactgagagtgc"
     misc_feature    6771..7631
                     /label=ampR
                     /note="color: #b1ff67"
ORIGIN
        1 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
       61 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      121 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      181 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      241 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      301 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      361 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      421 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      481 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      541 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      601 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      661 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      721 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      781 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      841 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      901 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      961 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1021 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1081 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1141 acaggaaaca gctatgacca tgattacgcc ggggcgcgcc attctagatt aaccctagaa
     1201 agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata
     1261 gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg
     1321 cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa
     1381 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
     1441 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
     1501 caactagaat gctagcatgg gcccagatct ccgaggggac ccgacaggcc cgatctgctt
     1561 tttgcttgta ctgtcgaccc tgtggaatga ccgtcgacct cgacggatcg ggagatctcg
     1621 accgagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga
     1681 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
     1741 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1801 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     1861 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     1921 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     1981 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
     2041 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
     2101 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
     2161 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
     2221 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
     2281 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2341 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2401 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2461 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2521 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     2581 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     2641 acggagtacc gggcgccgtc caggcacctc gattagttct cgatcgagct tttggagtac
     2701 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     2761 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     2821 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     2881 atttcaggtg tcgtgaggaa ttagcttgac caagcttggt accgagctcg gatccactag
     2941 taacggccgc cagtgtgctg gaattatcaa caagtttgta caaaaaagca ggctgctatg
     3001 accatgaaga tggacaaaaa gactatagtt tggtttagaa gagacctaag gattgaggat
     3061 aatcctgcat tagcagcagc tgctcacgaa ggatctgttt ttcctgtctt catttggtgt
     3121 cctgaagaag aaggacagtt ttatcctgga agagcttcaa gatggtggat gaaacaatca
     3181 cttgctcact tatctcaatc cttgaaggct cttggatctg acctcacttt aatcaaaacc
     3241 cacaacacga tttcagcgat cttggattgt atccgcgtta ccggtgctac aaaagtcgtc
     3301 tttaaccacc tctatgatcc tgtttcgtta gttcgggacc ataccgtaaa ggagaagctg
     3361 gtggaacgtg ggatctctgt gcaaagctac aatggagatc tattgtatga accgtgggag
     3421 atatactgcg aaaagggcaa accttttacg agtttcaatt cttactggaa gaaatgctta
     3481 gatatgtcga ttgaatccgt tatgcttcct cctccttggc ggttgatgcc aataactgca
     3541 gcggctgaag cgatttgggc gtgttcgatt gaagaactag ggctggagaa tgaggccgag
     3601 aaaccgagca atgcgttgtt aactagagct tggtctccag gatggagcaa tgctgataag
     3661 ttactaaatg agttcatcga gaagcagttg atagattatg caaagaacag caagaaagtt
     3721 gttgggaatt ctacttcact actttctccg tatctccatt tcggggaaat aagcgtcaga
     3781 cacgttttcc agtgtgcccg gatgaaacaa attatatggg caagagataa gaacagtgaa
     3841 ggagaagaaa gtgcagatct ttttcttagg ggaatcggtt taagagagta ttctcggtat
     3901 atatgtttca acttcccgtt tactcacgag caatcgttgt tgagtcatct tcggtttttc
     3961 ccttgggatg ctgatgttga taagttcaag gcctggagac aaggcaggac cggttatccg
     4021 ttggtggatg ccggaatgag agagctttgg gctaccggat ggatgcataa cagaataaga
     4081 gtgattgttt caagctttgc tgtgaagttt cttctccttc catggaaatg gggaatgaag
     4141 tatttctggg atacactttt ggatgctgat ttggaatgtg acatccttgg ctggcagtat
     4201 atctctggga gtatccccga tggccacgag cttgatcgct tggacaatcc cgcgttacaa
     4261 ggcgccaaat atgacccaga aggtgagtac ataaggcaat ggcttcccga gcttgcgaga
     4321 ttgccaactg aatggatcca tcatccatgg gacgctcctt taaccgtact caaagcttct
     4381 ggtgtggaac tcggaacaaa ctatgcgaaa cccattgtag acatcgacac agctcgtgag
     4441 ctactagcta aagctatttc aagaacccgt gaagcacaga tcatgatcgg agcagcagcc
     4501 cgggatccac cggtcgccac cgatggtgga ggaggtagtg gaggaggagg atcaaggatg
     4561 ttgtgtccac gtatgaaggg agatggggaa actatgacta atgactatgt agttcatgga
     4621 ccagcttctg tgcctcttgg ttatgtgcca cacccaagtt ctttgtcagg atctcttcca
     4681 ggaatgtctc gaggtaaatc aatgatcagc tccctcagta tcatgggggg aagcagtgga
     4741 cccccctatg accgagccca tgttacagga gcatcatcaa gtagttcttc aagcaccaaa
     4801 ggcacttact tccctgcaat tttgaaccct ccaccatccc cagccacaga gcgatcacat
     4861 tacactatgg aatttggata ttcttcaaac agtccttcca ctcataggtc atacagctac
     4921 aggccatata gctaccggca ctttgcaccc cccaccacac cctgcagcac agatgtttgt
     4981 gacagtgact atgctcctag tcggagaatg acctcagtgg caacagccaa gggctatacc
     5041 agtgacttga actatgattc agaacctgtg cccccacctc ccacaccccg aagccaatac
     5101 ttgtcagcag aggagaacta tgaaagctgc ccaccttctc catacacaga gaggagctat
     5161 tctcatcacc tctacccacc gccaccctct ccctgtacag actcctccag ttctggatcc
     5221 ggaggaagct ccgggagtgg ctcaggcagt tcaaagggta aaaagaagaa aaagaagtca
     5281 aagacaaagt gtgtaattat gggaagcgga gctactaact tcagcctgct gaagcaggct
     5341 ggagacgtgg aggagaaccc tggacctatg accgagtaca agcccacggt gcgcctcgcc
     5401 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5461 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5521 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5581 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5641 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5701 cttaacgccc gccccacgac ccctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5761 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5821 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5881 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgagtgccc
     5941 gaaggaccgc gcgacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     6001 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgatactag ttaaaagttt
     6061 tgttacttta tagaagaaat tttgagtttt tgtttttttt taataaataa ataaacataa
     6121 ataaattgtt tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat
     6181 ctattcaaat taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt
     6241 acgcatgatt atctttaacg tacgtcacaa tatgattatc tttctagggt taatctagta
     6301 tacgcgtatg cggccgctta attaatccat ggcgcctgat gcggtatttt ctccttacgc
     6361 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     6421 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6541 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6601 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6661 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6721 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6781 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6841 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6901 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6961 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7021 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7081 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7141 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7201 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7261 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7321 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7381 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7441 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7501 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7561 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7621 agcattggta actg
//
