LOCUS       sequence-354145-        6919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Sox2 fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  TITLE     McKinnon Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6919)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLPS-hSox2-gfp
FEATURES             Location/Qualifiers
     source          1..6919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    complement(538..571)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1689..1724
                     /codon_start=1
                     /product="6x(His-Asn) affinity tag"
                     /label=6xHN
                     /translation="HNHNHNHNHNHN"
     terminator      complement(1760..1854)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1875..2534)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2449..2468
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(2563..2596)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2812..3525
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             2812..3522
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(2851..2872)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3112..3131)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3459..3480
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3648..3769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3685..3704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3739..3758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3776..4231)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3913..3934)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4125..4144
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4258..4362
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4364..4721
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4364..4555
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4387..4407)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4572..4707
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4756..5550
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4756..5550
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4810..4829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5420..5439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5738..5757)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5782..5829
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6158..6746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6647..6666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcata
      541 acttcgtata gcatacatta tacgaagtta tcagtcgacg gtaccggaca tatgcccggg
      601 aattcggcac gaggcccggg ccccccaaag tcccggccgg gccgagggtc ggcggccgcc
      661 ggcgggccgg gcccgcgcac agcgcccgca tgtacaacat gatggagacg gagctgaagc
      721 cgccgggccc gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg gcggcggccg
      781 gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc ttcatggtgt
      841 ggtcccgcgg gcagcggcgc aagatggccc aggagaaccc caagatgcac aactcggaga
      901 tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga gacggagaag cggccgttca
      961 tcgacgaggc taagcggctg cgagcgctgc acatgaagga gcacccggat tataaatacc
     1021 ggccccggcg gaaaaccaag acgctcatga agaaggataa gtacacgctg cccggcgggc
     1081 tgctggcccc cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc ggcctgggcg
     1141 cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa cggctggagc aacggcagct
     1201 acagcatgat gcaggaccag ctgggctacc cgcagcaccc gggcctcaat gcgcacggcg
     1261 cagcgcagat gcagcccatg caccgctacg acgtgagcgc cctgcagtac aactccatga
     1321 ccagctcgca gacctacatg aacggctcgc ccacctacag catgtcctac tcgcagcagg
     1381 gcacccctgg catggctctt ggctccatgg gttcggtggt caagtccgag gccagctcca
     1441 gcccccctgt ggttacctct tcctcccact ccagggcgcc ctgccaggcc ggggacctcc
     1501 gggacatgat cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc gcccccagca
     1561 gacttcacat gtcccagcac taccagagcg gcccggtgcc cggcacggcc attaacggca
     1621 cactgcccct ctcacacatg ccaagctttc tagaccattc gtttggcgcg cgggcccagg
     1681 taagtggtca taatcataat cataatcata atcataatca caactagcct aggagatcct
     1741 ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     1801 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1861 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     1921 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1981 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     2041 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     2101 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     2161 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     2221 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     2281 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     2341 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     2401 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     2461 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     2521 atttttttct ccattttagc ttccttagct cctgaaagat ccataacttc gtatagcata
     2581 cattatacga agttatagat ccaatattat tgaagcattt atcagggtta ttgtctcatg
     2641 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2701 ccccgaaaag tgccacctga cgtggatctc gagctcaagc ttcgaattca gggtttcctt
     2761 gacaatatca tacttatcct gtcccttttt tttccacagc taccggtcgc gagcaagggc
     2821 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2881 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2941 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3001 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     3061 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3121 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3181 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3241 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     3301 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3361 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3421 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3481 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     3541 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3601 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3781 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3841 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3901 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3961 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4021 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4081 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4141 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4201 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4261 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4321 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4381 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4441 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4561 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4621 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4681 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4741 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4921 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5281 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5521 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5701 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5761 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5821 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5881 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5941 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6001 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6301 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6841 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     6901 agcccatata tggagttcc
//
