LOCUS       Exported                8044 bp ds-DNA     circular SYN 20-MAR-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8044
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    13..453
                     /label=F1 Ori
                     /note="color: #02abea; direction: RIGHT"
     misc_feature    585..607
                     /label=M13_Fw23
                     /note="color: #dd0907; direction: RIGHT"
     misc_feature    624..641
                     /label=Sce I site
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    648..669
                     /label=T7 Promoter
                     /note="color: #0000d3; direction: RIGHT"
     misc_feature    677..884
                     /label=miniTol2Left
                     /note="color: #996633; direction: RIGHT"
     misc_feature    902..921
                     /label=Untitled
                     /note="color: #a6acb3"
     misc_feature    958..1165
                     /label=Untitled
                     /note="color: #a6acb3"
     misc_feature    958..1165
                     /label=PM-02-MCS-D-USER 2
                     /note="color: #a6acb3"
     misc_feature    958..1165
                     /label=PM-02-MCS-D-USER 2
                     /note="color: #a6acb3"
     misc_feature    958..1165
                     /label=b-Globin Intron
                     /note="color: #008012; direction: RIGHT"
     misc_feature    complement(1153..1175)
                     /label=PR-04
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    1286..1288
                     /label=junction marker
                     /note="This marker was generated to preserve the boundary 
                     between two segments whose ends were joined."
                     /note="color: #a6acb3"
     misc_feature    1288..1294
                     /label=PM-460???-Sce-Tol2-LFNG-GFPnls-T2A-ERt-Cre-ERt-...
                     /note="color: #a6acb3"
     misc_feature    1294^1295
                     /label=eGFP
                     /note="color: #008012"
     misc_feature    1295..2176
                     /label=PM-460???-Sce-Tol2-LFNG-GFPnls-T2A-ERt-Cre-ERt-...
                     /note="color: #a6acb3"
     CDS             1295..2122
                     /codon_start=1
                     /label=GFP-nls
                     /note="color: #008012"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSRADPKKKRKVDPKKKRKVDPKKKRKVGSTGSR"
     misc_feature    1322..1324
                     /label=pGEMT_Sox9-Fse-Kpn-GFPnls-T2A-mERCreER-PA-nHA
                     /note="color: #a6acb3"
     misc_feature    2011^2012
                     /label=NLS
                     /note="color: #0000d3"
     misc_feature    2122^2123
                     /label=T2A
                     /note="color: #a6acb3"
     misc_feature    2175..2177
                     /label=junction marker
                     /note="This marker was generated to preserve the boundary 
                     between two segments whose ends were joined."
                     /note="color: #a6acb3"
     CDS             2180..5101
                     /codon_start=1
                     /label=ERT-Cre-ERT
                     /note="color: #f20884"
                     /translation="MAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPI
                     LYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLE
                     ILMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEE
                     FVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQR
                     LAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEE
                     TDQSHLATAGSTSSHSLQKYYITGEAEGFPATAVDNLLTVHQNLPALPVDATSDEVRKN
                     LMDMFRDRQAFSEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAV
                     KTIQQHLGQLNMLHRRPGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQ
                     VRSLMENSDRCQDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTL
                     VSTAGVEKALSLGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALE
                     GIFEATHRLIYGAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIV
                     MNYIRNLDSETGAMVRLLEDGDLEPSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQM
                     VSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHD
                     QVHLLECAWLEILMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSS
                     RFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAK
                     AGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHA
                     PTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATA"
     misc_feature    5111..5634
                     /label=Poly A
                     /note="color: #f20884; direction: RIGHT"
     misc_feature    complement(5637..5650)
                     /label=PR-36
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    5657..5806
                     /label=mTol2Right
                     /note="color: #996633; direction: RIGHT"
     misc_feature    complement(5832..5851)
                     /label=T3 Promoter
                     /note="color: #0000d3; direction: LEFT"
     misc_feature    complement(5858..5875)
                     /label=Sce I Site
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    complement(5908..5930)
                     /label=M13_Rv23
                     /note="color: #dd0907; direction: LEFT"
     misc_feature    complement(6279..6961)
                     /label=ColE1 origin
                     /note="color: #008012; direction: LEFT"
     misc_feature    complement(7056..7916)
                     /label=Amp Res
                     /note="color: #0000d3; direction: LEFT"
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgtagggat aacagggtaa tgcgcgcgta atacgactca
      661 ctatagggcg aattggcaga ggtgtaaagt acttgagtaa ttttacttga ttactgtact
      721 taagtattat ttttggggat ttttacttta cttgagtaca attaaaaatc aatactttta
      781 cttttactta attacatttt tttagaaaaa aaagtacttt ttactcctta caattttatt
      841 tacagtcaaa aagtacttat tttttggaga tcacttgggc ccggctcgag catgcacgcg
      901 ttagcggccg ctaggtacca tgaattcgct gagggcgttt aaaatcctca gcaattcgaa
      961 ttttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1021 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1081 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1141 cccagttcaa ttacagctct taaaattccg ccactagagc ctctgctaac catgttcatg
     1201 ccttcttctc tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1261 ttggcaaaga attattccgc tagcgctcgc caccatggtg agcaagggcg aggagctgtt
     1321 caccggggtg gtacccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1381 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1441 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1501 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1561 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1621 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1681 cgacttcaag gaggacggca acatcctggg gcacaagcta gagtacaact acaacagcca
     1741 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1801 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1861 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1921 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1981 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag ctgatccaaa
     2041 aaagaagaga aaggtagatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa
     2101 ggtaggatcc accggatcta gagagggcag aggaagtctt ctaacatgcg gtgacgtgga
     2161 ggagaatccc ggccctgtca tggccggcga tatgagagcc gctaacctgt ggccctcccc
     2221 actgatgatc aagagaagca agaagaactc cctggccctg agcctgaccg ctgatcagat
     2281 ggtgtccgcc ctgctggacg ccgagccccc tatcctgtac agcgagtacg accccaccag
     2341 gcccttcagc gaggccagca tgatgggact gctgacaaac ctggccgatc gcgaactggt
     2401 gcatatgatc aattgggcca agcgcgtgcc cggattcgtg gacctgacac tgcacgacca
     2461 ggtgcacctg ctggaatgcg cttggctgga aattctgatg atcggcctcg tgtggcggag
     2521 catggaacat cctgtgaagc tgctgttcgc ccccaatctg ctgctggacc gcaaccaggg
     2581 caaatgcgtg gaagggatgg tcgaaatctt tgatatgctg ctcgccacca gcagcaggtt
     2641 cagaatgatg aacctccagg gcgaagagtt cgtctgcctg aagtccatca tcctgctgaa
     2701 cagcggcgtg tacaccttcc tgtcctctac cctgaagtcc ctcgaggaaa aggatcacat
     2761 ccacagggtg ctggataaga tcaccgatac cctgatccat ctcatggcta aggccggact
     2821 cacactccag cagcagcatc agagactcgc tcagctgctg ctgatcctga gccacatccg
     2881 ccacatgtcc aacaagggaa tggaacacct ctactctatg aagtgtaaaa acgtcgtccc
     2941 cctgtacgat ctgctgctcg aggccgccga tgcccacaga ctgcacgccc ctacaagcag
     3001 aggcggcgct agcgtggaag agacagacca gtcccacctg gccaccgccg gcagcacaag
     3061 cagccacagc ctgcagaagt actatatcac cggcgaggcc gagggcttcc ccgctacagc
     3121 tgtcgacaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     3181 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     3241 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     3301 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
     3361 gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg
     3421 tcggcccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
     3481 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
     3541 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
     3601 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
     3661 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
     3721 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
     3781 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
     3841 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
     3901 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg atgactccgg
     3961 tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc
     4021 ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat
     4081 tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga
     4141 agatggcgat ctcgagccat ctgctggaga catgagagct gccaaccttt ggccaagccc
     4201 gctcatgatc aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat
     4261 ggtcagtgcc ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag
     4321 acccttcagt gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt
     4381 tcacatgatc aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca
     4441 ggtccacctt ctagaatgtg cctggctaga gatcctgatg attggtctcg tctggcgctc
     4501 catggagcac ccagtgaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg
     4561 aaaatgtgta gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt
     4621 ccgcatgatg aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa
     4681 ttctggagtg tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat
     4741 ccaccgagtc ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct
     4801 gaccctgcag cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag
     4861 gcacatgagt aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc
     4921 cctctatgac ctgctgctgg aggcggcgga cgcccaccgc ctacatgcgc ccactagccg
     4981 tggaggggca tccgtggagg agacggacca aagccacttg gccactgcgg gctctacttc
     5041 atcgcattcc ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagc
     5101 ttaagatatc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     5161 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     5221 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     5281 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     5341 aaaacatcag aatgagcatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     5401 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     5461 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     5521 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     5581 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa gatccctcga
     5641 ccagcggccg gccgctaata ctcaagtaca attttaatgg agtacttttt tacttttact
     5701 caagtaagat tctagccaga tacttttact tttaattgag taaaattttc cctaagtact
     5761 tgtactttca cttgagtaaa atttttgagt actttttaca cctctgccgc ggtggagctc
     5821 cagcttttgt tccctttagt gagggttaat tgcgcgcatt accctgttat ccctacgcgc
     5881 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5941 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6001 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7921 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8041 ccac
//
