pDG172

LOCUS       pDG172                8509 bp DNA     circular SYN 10-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    13..38
                     /label=SleepingBeauty-ITR-Li
     misc_feature    209..240
                     /label=SleepingBeauty-ITR_Li
     primer_bind     276..317
                     /label=oMH09-pWS252-EF1a-
                     /note="AGGGGTCCGCTATCTACGGGAAAGTGATGTCGTGTAC 同源臂"
     misc_feature    298..1364
                     /label=P_EF1a
     intron          417..1355
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(1335..1365)
                     /label=Fragment.REV
     intron          1373..1505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    1373..1505
                     /label=CI
     regulatory      1547..1556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1556..2491
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2501..3523
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
                     VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
                     PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
                     IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
                     ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
                     LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
                     D"
     CDS             3536..4471
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     primer_bind     complement(4486..4502)
                     /label=Fragment.REV
     polyA_signal    4493..4684
                     /label=SV40 late polyA
     promoter        4826..5325
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5333..5932
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     terminator      5953..6180
                     /label=bGH PA
     misc_feature    6264..6292
                     /label=SleepingBeauty-ITR_Ri
     misc_feature    6458..6489
                     /label=SleepingBeauty-ITR_Ro
     rep_origin      complement(6659..7287)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(7436..8296)
                     /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"
     promoter        complement(8297..8401)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(8338..8366)
                     /label=Amp prom
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct actcgacggg
      301 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      361 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      421 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
      481 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
      541 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
      601 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
      661 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
      721 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
      781 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
      841 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
      901 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
      961 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
     1021 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     1081 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     1141 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1201 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     1261 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1321 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgacctgca gggtaagtat
     1381 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     1441 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     1501 cacagggtac cgagctcgga tccactagtc cagtgtggtg gaattcgcca ccatggctgg
     1561 agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa
     1621 cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc
     1681 ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg
     1741 cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt
     1801 gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct
     1861 agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga agctactgtt
     1921 tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat
     1981 cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga
     2041 gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag
     2101 caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga
     2161 cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct
     2221 ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca
     2281 tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc tggaggcggc
     2341 ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacgga
     2401 ccaaagccac ttggccactg cgggctctac ttcatcgcat tccttgcaaa agtattacat
     2461 cacgggggag gcagagggtt tccctgccac agctgtcgac aatttactga ccgtacacca
     2521 aaatttgcct gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga acctgatgga
     2581 catgttcagg gatcgccagg cgttttctga gcatacctgg aaaatgcttc tgtccgtttg
     2641 ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa tggtttcccg cagaacctga
     2701 agatgttcgc gattatcttc tatatcttca ggcgcgcggt ctggcagtaa aaactatcca
     2761 gcaacatttg ggccagctaa acatgcttca tcgtcggtcc gggctgccac gaccaagtga
     2821 cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga
     2881 acgtgcaaaa caggctctag cgttcgaacg cactgatttc gaccaggttc gttcactcat
     2941 ggaaaatagc gatcgctgcc aggatatacg taatctggca tttctgggga ttgcttataa
     3001 caccctgtta cgtatagccg aaattgccag gatcagggtt aaagatatct cacgtactga
     3061 cggtgggaga atgttaatcc atattggcag aacgaaaacg ctggttagca ccgcaggtgt
     3121 agagaaggca cttagcctgg gggtaactaa actggtcgag cgatggattt ccgtctctgg
     3181 tgtagctgat gatccgaata actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc
     3241 gccatctgcc accagccagc tatcaactcg cgccctggaa gggatttttg aagcaactca
     3301 tcgattgatt tacggcgcta aggatgactc tggtcagaga tacctggcct ggtctggaca
     3361 cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa taccggagat
     3421 catgcaagct ggtggctgga ccaatgtaaa tattgtcatg aactatatcc gtaacctgga
     3481 tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gatctcgagc catctgctgg
     3541 agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa
     3601 cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc
     3661 ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg
     3721 cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt
     3781 gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct
     3841 agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga agctactgtt
     3901 tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat
     3961 cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga
     4021 gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag
     4081 caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga
     4141 cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct
     4201 ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca
     4261 tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc tggaggcggc
     4321 ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacgga
     4381 ccaaagccac ttggccactg cgggctctac ttcatcgcat tccttgcaaa agtattacat
     4441 cacgggggag gcagagggtt tccctgccac agcttgatag cggcagacgc gtcagacatg
     4501 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4561 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4621 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4681 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg
     4741 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4801 cttgtctgtt ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     4861 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     4921 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     4981 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     5041 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     5101 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     5161 ttctgggctc agaggctggg aatgggtggg tccgggggcg ggctcagggg cgggctcagg
     5221 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     5281 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg caatgaccga
     5341 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5401 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5461 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5521 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5581 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5641 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5701 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5761 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5821 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5881 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gatagagctc
     5941 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     6001 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     6061 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     6121 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6181 cttctgaggc ggaaagaacc agctgggcct actagctact cgggacccct taccgaaaca
     6241 tcgccgcatt ctgcagagga gtcgagtgta tgtaaacttc tgacccactg ggaatgtgat
     6301 gaaagaaata aaagctgaaa tgaatcattc tctctactat tattctgata tttcacattc
     6361 ttaaaataaa gtggtgatcc taactgacct aagacaggga atttttacta ggattaaatg
     6421 tcaggaattg tgaaaaagtg agtttaaatg tatttggcta aggtgtatgt aaacttccga
     6481 cttcaactgt atagggatcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8401 gcacatttcc ccgaaaagtg ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg
     8461 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaatt
//