LOCUS       sequence-384896-        6745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tagging human SOX2 gene with P2A-EGFP at the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Sidhaye J, Trepte P, Sepke N, Novatchkova M, Schutzbier M, 
            Durnberger G, Mechtler K, Knoblich JA
  TITLE     Integrated transcriptome and proteome analysis reveals 
            posttranscriptional regulation of ribosomal genes in human brain 
            organoids.
  JOURNAL   Elife. 2023 Mar 29;12:e85135. doi: 10.7554/eLife.85135.
  PUBMED    36989136
REFERENCE   2  (bases 1 to 6745)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2023-03-29"; volume: "12:e85135. doi"; pages: " 
            10.7554/eLife.85135"
COMMENT     Sequence Label: pHDR_sox2HA_P2A-GFP-pA_HA_G2913A
FEATURES             Location/Qualifiers
     source          1..6745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(261..365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(391..846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(528..549)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     740..759
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     973..995
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     987..1004
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     988..1004
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1014..1033
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1014..1032
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1130..1195
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1272..1844)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     CDS             3121..3177
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(3184..3186,3187..3189,3190..3903)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3184..3186,3187..3189,3190..3900)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3229..3250)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3490..3509)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3837..3858
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4984..5000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4984..5000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4997..5019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5008..5024
                     /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)."
     promoter        complement(join(5032..5038,5039..5056,5057..5062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5077..5098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5215..5232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5386..5974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5466..5485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6145..6745,1..191,192..260))
                     /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"
ORIGIN
        1 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
       61 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
      121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
      181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
      241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
      301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      361 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
      421 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
      481 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
      541 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
      601 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
      661 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
      721 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
      781 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
      841 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      901 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      961 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     1021 actcactata gggcgaattg ggtaccgggc cccccctcga ggtccccctc gactctagtg
     1081 gatctgcatt ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac
     1141 aaacaattag aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag
     1201 agctttaaat ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct
     1261 tcacaaagag atcgcctgac acgatttcct gcacaggctt gagccatata ctcatacatc
     1321 gcatcttggc cacgttttcc acgggtttca aaattaatct caagttctac gcttaacgct
     1381 ttcgcctgtt cccagttatt aatatattca acgctagaac tcccctcagc gaagggaagg
     1441 ctgagcacta cacgcgaagc accatcaccg aaccttttga taaactcttc cgttccgact
     1501 tgctccatca acggttcagt gagacttaaa cctaactctt tcttaatagt ttcggcatta
     1561 tccactttta gtgcgagaac cttcgtcagt cctggatacg tcactttgac cacgcctcca
     1621 gcttttccag agagcgggtt ttcattatct acagagtatc ccgcagcgtc gtatttattg
     1681 tcggtactat aaaacccttt ccaatcatcg tcataatttc cttgtgtacc agattttggc
     1741 ttttgtatac ctttttgaat ggaatctaca taaccaggtt tagtcccgtg gtacgaagaa
     1801 aagttttcca tcacaaaaga tttagaagaa tcaacaacat catcaggatc catggcgagg
     1861 acctgcaggg tcgctcggtg ttcgaggcca cacgcgtcac cttaatatgc gaagtggacc
     1921 tgggaccgcg ccgccccgac tgcatctgcg tgttcgaatt cgccaatgac aagacgctgg
     1981 gcggggtttg ctcgacattg ggtggaaaca ttccaggcct gggtggagag gctttttgct
     2041 tcctcttgca aaaccacact gctcgacatt gggtggaaac attccaggcc tgggtggaga
     2101 ggctttttgc ttcctcttga aaaccacact gcgacggtat cgataagctt gatatcgtcg
     2161 accatgatgg agacggagct gaagccgccg ggcccgcagc aaacttcggg gggcggcggc
     2221 ggcaactcca ccgcggcggc ggccggcggc aaccagaaaa acagcccgga ccgcgtcaag
     2281 cggcccatga atgccttcat ggtgtggtcc cgcgggcagc ggcgcaagat ggcccaggag
     2341 aaccccaaga tgcacaactc ggagatcagc aagcgcctgg gcgccgagtg gaaacttttg
     2401 tcggagacgg agaagcggcc gttcatcgac gaggctaagc ggctgcgagc gctgcacatg
     2461 aaggagcacc cggattataa ataccggccc cggcggaaaa ccaagacgct catgaagaag
     2521 gataagtaca cgctgcccgg cgggctgctg gcccccggcg gcaatagcat ggcgagcggg
     2581 gtcggggtgg gcgccggcct gggcgcgggc gtgaaccagc gcatggacag ttacgcgcac
     2641 atgaacggct ggagcaacgg cagctacagc atgatgcagg accagctggg ctacccgcag
     2701 cacccgggcc tcaatgcgca cggcgcagcg cagatgcagc ccatgcaccg ctacgacgtg
     2761 agcgccctgc agtacaactc catgaccagc tcgcagacct acatgaacgg ctcgcccacc
     2821 tacagcatgt cctactcgca gcagggcacc cctggcatgg ctcttggctc catgggttcg
     2881 gtggtcaagt ccgaggccag ctccagcccc cctgtggtta cctcttcctc ccactccagg
     2941 gcgccctgcc aggccgggga cctccgggac atgatcagca tgtatctccc cggcgccgag
     3001 gtgccggaac ccgccgcccc cagcagactt cacatgtccc agcactacca gagcggcccg
     3061 gtgcccggca cagccattaa cggcacactg cccctctcac acatggtcga cggaagcgga
     3121 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctctc
     3181 gacatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3241 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3301 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3361 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3421 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3481 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3541 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3601 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3661 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3721 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3781 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3841 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3901 taagggccgg acagcgaact ggagggggga gaaattttca aagaaaaacg agggaaatgg
     3961 gaggggtgca aaagaggaga gtaagaaaca gcatggagaa aacccggtac gctcaaaaag
     4021 aaaaaggaaa aaaaaaaatc ccatcaccca cagcaaatga cagctgcaaa agagaacacc
     4081 aatcccatcc acactcacgc aaaaaccgcg atgccgacaa gaaaactttt atgagagaga
     4141 tcctggactt ctttttgggg gactattttt gtacagagaa aacctgggga gggtggggag
     4201 ggcgggggaa tggaccttgt atagatctgg aggaaagaaa gctacgaaaa actttttaaa
     4261 agttctagtg gtacggtagg agctttgcag gaagtttgca aaagtcttta ccaataatat
     4321 ttagagctag tctccaagcg acgaaaaaaa tgttttaata tttgcaagca acttttgtac
     4381 agtatttatc gagataaaca tggcaatcaa aatgtccatt gtttataagc tgagaatttg
     4441 ccaatatttt tcaaggagag gcttcttgct gaattttgat tctgcagctg aaatttagga
     4501 cagttgcaaa cgtgaaaaga agaaaattat tcaaatttgg acattttaat tgtttaaaaa
     4561 ttgtacaaaa ggaaaaaatt agaataagta ctggcgaacc atctctgtgg tcttgtttaa
     4621 aaagggcaaa agttttagac tgtactaaat tttataactt actgttaaaa gcaaaaatgg
     4681 ccatgcaggt tgacaccgtt ggtaatttat aatagctttt gttcgatccc aactttccat
     4741 tttgttcaga taaaaaaaac catgaaatta ctgtgtttga aatattttct tatggtttgt
     4801 aatatttctg taaatttatt gtgatatttt aaggttttcc cccctttatt ttccgtagtt
     4861 gtattttaaa agattcggct ctgtattatt tgaatcagtc tgcgtttaaa cccgccaccg
     4921 cggtggagct ccagcttttg ttccctttta gtgagggtta attgcgcgct tggcgtaatc
     4981 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5041 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5101 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5161 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6001 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6721 atagtgtatg cggcgaccga gttgc
//
