LOCUS       hPGK_mNG_T2A_mCh        4362 bp DNA     circular SYN 08-JAN-2025
DEFINITION  .
ACCESSION   VB241016-1888hha
VERSION     .
KEYWORDS    VB241016-1888hha.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4362
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     promoter        22..526
                     /label=hPGK
                     /note="/Application Notes=Medium-strength promoter."
                     /note="/References=PLoS One. 4:e5046(2009)"
                     /note="/Subtype=Mammalian ubiquitous promoter"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Medium-strength promoter."
                     /note="/description=Human phosphoglycerate kinase 1 
                     promoter"
                     /note="/name=hPGK"
                     /note="/official_designation=hPGK promoter"
                     /note="/uuid=a105ee8f48814e769c9a41c38cfff41e"
                     /note="color: #800c0c; direction: RIGHT"
     misc_feature    551..556
                     /label=NheI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name=NheI"
                     /note="/official_designation=BamHI"
                     /note="/uuid=efd7b4e467c64c70ae8bfad8c16a27cb"
                     /note="color: #0c5d90; direction: RIGHT"
     misc_feature    557..562
                     /label=Kozak
                     /note="/Application Notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/Full Name=Kozak"
                     /note="/References=Nucleic Acids Res. 9:5233 (1981)"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/full_name=Kozak"
                     /note="/name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
                     /note="color: #f58600; direction: RIGHT"
     misc_feature    563..1270
                     /label={mNeoGreen}
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name={mNeoGreen}"
                     /note="/official_designation={mNeoGreen}"
                     /note="/uuid=fb274bd8f4414b8db7187c16b1ed5b32"
                     /note="color: #5566f5; direction: RIGHT"
     misc_feature    1271..1276
                     /label=EcoRI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name=EcoRI"
                     /note="/official_designation=EcoRI"
                     /note="/uuid=18b38e5fe9db4734be3e7bd8a74a83da"
                     /note="color: #ef6cdf; direction: RIGHT"
     misc_feature    1277..1339
                     /label=T2A
                     /label=nonstandard type:  Linker
                     /note="/Application Notes=Causes co-translational cleavage 
                     of theencoded polypeptide. Multiple proteins can be made 
                     from a polycistronic transcript containing multiple ORFs 
                     separated by 2A. P2A and T2A have higher cleavage 
                     efficiency compared to other 2As."
                     /note="/Full Name=Thosea asigna virus 2A peptide"
                     /note="/References=PLoS One. 6:e18556(2011)"
                     /note="/Subtype=Read-through linker"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Causes co-translational cleavage 
                     of theencoded polypeptide. Multiple proteins can be made 
                     from a polycistronic transcript containing multiple ORFs 
                     separated by 2A. P2A and T2A have higher cleavage 
                     efficiency compared to other 2As."
                     /note="/description=Self-cleaving 2A peptide from Thosea 
                     asignavirus"
                     /note="/full_name=Thosea asigna virus 2A peptide"
                     /note="/name=T2A"
                     /note="/official_designation=T2A"
                     /note="/uuid=995274d0c39f4cbdbab66fb78aa938d6"
                     /note="color: #c54b7c; direction: RIGHT"
     misc_feature    1340..1345
                     /label=BamHI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name=BamHI"
                     /note="/official_designation=BamHI"
                     /note="/uuid=2e24d26a052f4a8f9539513b7d1f974d"
                     /note="color: #d05c0a; direction: RIGHT"
     misc_feature    1346..2056
                     /label={mCh}
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name={mCh}"
                     /note="/official_designation={mCh}"
                     /note="/uuid=3e0ba29352f64ebc96467baf4bfb9961"
                     /note="color: #fd3434; direction: RIGHT"
     misc_feature    2060..2065
                     /label=KpnI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name=KpnI"
                     /note="/official_designation=KpnI"
                     /note="/uuid=0ca3b9c82c704e30800d68808aa4b838"
                     /note="color: #46c6ef; direction: RIGHT"
     polyA_signal    2110..2331
                     /label=SV40 late pA
                     /note="/Application Notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/References=Mol. Cell. Biol.9:4248(1989)"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Simian virus 40 late polyadenylation 
                     signal"
                     /note="/name=SV40 late pA"
                     /note="/official_designation=SV40 late pA"
                     /note="/uuid=d23bf533e0924dfcb932fc3ddd77e791"
                     /note="color: #eda481; direction: RIGHT"
     rep_origin      complement(2527..3115)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/Application Notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/Full Name=pUC origin of replication"
                     /note="/References=Mol Microbiol. 6:3385 (1992)"
                     /note="/Subtype=E. coli replication_origin"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/name=pUC ori"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=41c627938a364ebfa136b55444385c5b"
                     /note="color: #da3bb5"
     CDS             complement(3286..4146)
                     /label=Ampicillin
                     /note="/Application Notes=Allows E. coli to be resistant 
                     toampicillin."
                     /note="/Full Name=Ampicillin resistance gene"
                     /note="/References=Proc Natl Acad Sci U S A. 75: 3737 
                     (1978)"
                     /note="/Selection Drug=Ampicillin "
                     /note="/Subtype=E. coli selection marker"
                     /note="/Usage=Positive selection"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows E. coli to be resistant 
                     toampicillin."
                     /note="/description=AmpiciIIin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/name=Ampicillin"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=0bc7d58885794858b29e98fe171ff1d5"
                     /note="color: #e4b930; direction: LEFT"
ORIGIN
        1 caactttgta tagaaaagtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag
       61 ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc
      121 acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc
      181 ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc
      241 cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag
      301 ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg
      361 cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg
      421 tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg
      481 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggcaag tttgtacaaa
      541 aaagcaggct gctagcgcca ccatggtgag taagggggag gaagataaca tggcctcact
      601 tccagcgact cacgaacttc acatttttgg gtccatcaat ggcgtggatt tcgatatggt
      661 agggcaaggg actggcaacc ctaacgacgg ttacgaagaa ctcaatctga aatcaacgaa
      721 aggggatttg caattttccc catggatact cgtcccccat ataggctatg gattccacca
      781 atatctgccg tatccagacg gaatgtctcc gtttcaagcc gcaatggtag acggtagcgg
      841 ttaccaggtc caccggacga tgcaattcga agatggggcc tccctgacag taaattatag
      901 atatacctat gaagggagtc acataaaggg agaagctcag gtcaaaggaa cgggtttccc
      961 ggctgacgga ccggtgatga ccaattcact gaccgccgcc gattggtgcc gctctaaaaa
     1021 aacttaccca aatgataaga ctatcatttc aacttttaaa tggagttaca ctacggggaa
     1081 tggtaagagg tacaggtcaa cagccaggac aacttacaca tttgccaagc caatggcggc
     1141 gaactatttg aaaaatcagc caatgtacgt tttccgaaaa accgaattga aacattctaa
     1201 gaccgagctt aatttcaagg aatggcaaaa agcatttacg gatgtgatgg ggatggatga
     1261 gctttacaag gaattcggaa gcggagaggg caggggaagt cttctaacat gcggggacgt
     1321 ggaggaaaat cccggccccg gatccatggt ctccaaagga gaggaagaca atatggccat
     1381 tatcaaagag tttatgcgat ttaaagtgca catggaggga agcgtcaacg ggcatgagtt
     1441 tgagattgag ggggaaggag agggaagacc ttatgaaggg acccagactg ccaaactgaa
     1501 agtcaccaag ggcggacccc tgccatttgc ttgggacatt ctgagcccac aattcatgta
     1561 tgggtccaaa gcctatgtta aacacccggc agacatacct gactacctca agctctcctt
     1621 tcctgagggg tttaaatggg agagggtgat gaactttgaa gatggaggcg tcgtgacagt
     1681 aactcaggat agcagcctgc aagacgggga gtttatttac aaggttaaat tgcgcggaac
     1741 caactttccc agcgatggac ccgtgatgca gaagaaaaca atgggctggg aagcatcatc
     1801 cgaaagaatg tatcccgaag atggcgcgtt gaagggggaa atcaaacaaa ggctcaagtt
     1861 gaaagatgga gggcactatg acgctgaggt caaaaccacg tacaaggcta aaaaaccggt
     1921 acaactgccc ggagcatata atgttaatat taagctggac atcaccagcc acaatgagga
     1981 ttacacaata gtggaacagt atgaaagagc cgagggaaga cactcaactg ggggcatgga
     2041 tgaactgtat aaataataag gtaccaccca gctttcttgt acaaagtggt gatggccggc
     2101 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2161 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2221 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2281 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt agcggccgcg
     2341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctctcttc
     2701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4261 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4321 taggcgtatc acgaggccct ttcgtcggcg cgccgcggcc gc
//
