LOCUS       sequence-9927-ex        8448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8448)
  AUTHORS   Dann CT, Alvarado AL, Molyneux LA, Denard BS, Garbers DL, Porteus MH
  TITLE     Spermatogonial stem cell self-renewal requires OCT4, a factor 
            downregulated during retinoic acid-induced differentiation.
  JOURNAL   Stem Cells. 2008 Nov . 26(11):2928-37.
  PUBMED    18719224
REFERENCE   2  (bases 1 to 8448)
  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: "Stem Cells.
            2008 Nov . 26(11):2928-37."
COMMENT     Sequence Label: pLSG (stra8)
FEATURES             Location/Qualifiers
     source          1..8448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..2935
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2854..2874
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2952..2968
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3010..3043)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4503..4512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4509..4511,4512..4514,4515..5228)
                     /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(4509..4511,4512..4514,4515..5225)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4554..4575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4815..4834)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5162..5183
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5248..5281)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5337..5925
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5390..5410)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5808..5819)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5928..5944)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6454..6634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6696..7284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6776..6795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7455..8246,8247..8315))
                     /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"
     primer_bind     8078..8097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8316..8420)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg ccgccatctc
     2641 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2701 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2761 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2821 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2881 gtaaagtaat tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgttaacgc
     2941 gcggtgaccc tcgaggtcga cggtatcgat aagctcgctt cacgagattc cagcaggtcg
     3001 agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttccaagct
     3061 taagcggccc gcgggcccct agtcttgcct ccaagggggt aaggtgtagc tgctctttgg
     3121 acctggtaca agcctttgag agaggccctg ctgccattcc tgtcctcagc actcttttct
     3181 gtatcggtct ggcaattcac tacagtgtct tacaaagctc cagctccttt agccagccat
     3241 atccctgata gtttagaggg gagctggtaa ctgggtcccc actggtcccc actcctgtac
     3301 cttaagcatc agatctgttt tctgtcagga gcaagagcct catttctaag ctcaggttag
     3361 ccaccaactt catttttttt ttctttctgg gctacctgat tgctgcgact acagctatga
     3421 tccaccattc ccactgaaca gttttcattt tgtttaaaat ccacaatcta ccaaaaccat
     3481 ttgaaagatg acaccaggac tcgctagagt caaatttaag gtaccctgtt gcatacttta
     3541 gaaaggggct cctttctagc cagtcacagt caccgtgagg tagatcccgg attggaatcc
     3601 aacttctggt gctaccaaag tcgtggctgc agctgctacc tcaagtgacc tccgtttaac
     3661 ctctgtggag gggcgggggc gaggatggtg aagcagctca cctcaggtca gactgcagga
     3721 ggcaccctcc ctcctattca caggctgagc ctctcccctt ccaaggggct tttgcttgct
     3781 ttcctttctc tacaagagtg cgctcctagt gtgccagttt gatatggtgg cgctcgtagt
     3841 gtgccaaact gatgtggtgg tgtcaggacc attgcagata tctcaagatg cgagggaaaa
     3901 aagtggttta aagataatag cacatctgat ttgaggcgga aaatgagttg tagtccctca
     3961 acctggagaa aacttgttgc ttactacaaa caactcaaca catcctctct ctctctctct
     4021 tttcttctct gcattttaag tccaccttta aggttctctt ctctgctttt ttagttggga
     4081 atcccctatt cccctctcct attttgtacc tattccctct cacatcttca tttttctctt
     4141 tcttttcctt gaaacagggg actgctactg ggaccttgaa gatggctcct ctatatctca
     4201 agagaaagtt ataggtggca ttgccctggt tgaggggtgt aagaactggc gctagccgcc
     4261 tggatggggt gaaaaggtca tcttgctcct tccacaccct cttgcaacct gtggcaagtt
     4321 gttacaatgt tttcaccaat gtccacgctc cccattggcg ccccaccatg catccccatt
     4381 ggtcatggtg gcagtgacag ggctgtgatt ggttcgcagc ctggggtacc aggtcagttt
     4441 tttacctgag gcaagagcct ctcttcttat tctgcgacgg gcagtcgaat tcgctaccgg
     4501 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4561 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4621 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4681 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4741 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4801 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4861 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4921 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4981 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     5041 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     5101 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     5161 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     5221 acaagtagga attcgtcgag ggacctaata acttcgtata gcatacatta tacgaagtta
     5281 tacatgttta agggttccgg ttccactagg tacaattcga tatcaagctt atcgataatc
     5341 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5401 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5461 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5521 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5581 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5641 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5701 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5761 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5821 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5881 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     5941 ctcgatcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6001 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6061 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6121 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6181 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     6241 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     6301 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     6361 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     6421 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     6481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6601 agaccctttt agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag
     6661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7201 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7261 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7321 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7381 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7441 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7501 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7561 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7621 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7681 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7741 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7801 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7861 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7921 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7981 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8041 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8101 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8161 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8221 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8281 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8341 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8401 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
