LOCUS       sequence-36604-e        6403 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 6403)
  AUTHORS   Pasque V, Gillich A, Garrett N, Gurdon JB
  TITLE     Histone variant macroH2A confers resistance to nuclear 
            reprogramming.
  JOURNAL   EMBO J. 2011 May 6;30(12):2373-87. doi: 10.1038/emboj.2011.144.
  PUBMED    21552206
REFERENCE   2  (bases 1 to 6403)
  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"; doi: 
            "10.1038/emboj.2011"; journalName: "EMBO J"; date: "2011-05-6- 6"; 
            volume: "30"; issue: "12"; pages: "2373-87"
COMMENT     Sequence Label: pSUPER retro puro macroH2A1 shRNA
FEATURES             Location/Qualifiers
     source          1..6403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..24)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9..25)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(43..459)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(59..75)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(91..113)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    526..867
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             931..1447
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1647..1666)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1794..1816)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2010..2029)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2129..2151
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2189..2207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2275..2379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2380..2448,2449..3240)
                     /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     complement(2598..2617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3411..3999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3900..3919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4153..4170
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4287..4308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4323..4328,4329..4346,4347..4353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4361..4377
                     /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)."
     primer_bind     4366..4388
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4385..4401
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4385..4401
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(4994..5593)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5077..5097)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5574..5593
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5618..6117)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5666..5685)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6059..6082
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        6128..6342
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     6263..6282
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 agcttatcga taccgtcgac ctcgagagat cgatctaatt ccggcgccta gagaaggagt
       61 gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga
      121 cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct tgacccaggg
      181 aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt ctgggtgtcc
      241 atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc ggttaaaggt
      301 gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga aggtaaccca
      361 acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat ctttccagtg
      421 acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag acaaatacag
      481 aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca gacaagacgc
      541 gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt
      601 ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg actcccttcc
      661 tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag ggttgcggcc
      721 gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
      781 ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa agacagagac
      841 agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg gtccctgggc
      901 aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg ggcagtcaat
      961 caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca aacagcaaga
     1021 ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga
     1081 gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga
     1141 gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac
     1201 cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat ctggggacca
     1261 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
     1321 tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc agttatgtat
     1381 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc
     1441 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct atggctcgta
     1501 ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga ccactgatat
     1561 cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg ccggcatcac
     1621 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     1681 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     1741 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc cgtcgtttta
     1801 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1861 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1921 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1981 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2041 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2101 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2161 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2221 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2281 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2341 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2401 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2461 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2521 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2581 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2641 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2701 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2761 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2821 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2881 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2941 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3001 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3061 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3121 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3181 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3241 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3301 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3361 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3421 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3481 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3541 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3601 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3661 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3721 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3781 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3841 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3901 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3961 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4021 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4081 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4141 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4201 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4261 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4321 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4381 cacacaggaa acagctatga ccatgattac gaattaattc tccccagctc ttctgcgctg
     4441 tccataatga actatttcag gagtattgga gtatgtggat tcaaaacagt ggctaattga
     4501 gtcaggatta cttaatattc gactcctacc ctcagaatgt tgttctccaa cttcaagaaa
     4561 ctgttaccca tgaaagaccc ccgctgacgg gtagtcaatc actcagagga gaccctccca
     4621 aggaacagcg agaccacaag tcggatgcaa ctgcaagagg gtttattgga tacacgggta
     4681 cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt gggctctttt
     4741 attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt
     4801 caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct
     4861 ctagcttgcc aaacctacag gtggggtctt tcattccccc ctttttctgg agactaaata
     4921 aaatctttta ttttatcgat gcatggggtc gtgcgctcct ttcggtcggg cgctgcgggt
     4981 cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     5041 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     5101 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     5161 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
     5221 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     5281 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     5341 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     5401 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     5461 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
     5521 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
     5581 tgtactcggt catggtaagc tagcttgggc tgcaggtcga aaggcccgga gatgaggaag
     5641 aggagaacag cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg cctccggagg
     5701 accttcgggc gcccgccccg cccctgagcc cgcccctgag cccgcccccg gacccacccc
     5761 ttcccagcct ctgagcccag aaagcgaagg agcaaagctg ctattggccg ctgccccaaa
     5821 ggcctacccg cttccattgc tcagcggtgc tgtccatctg cacgagacta gtgagacgtg
     5881 ctacttccat ttgtcacgtc ctgcacgacg cgagctgcgg ggcggggggg aacttcctga
     5941 ctaggggagg agtagaaggt ggcgcgaagg ggccaccaaa gaacggagcc ggttggcgcc
     6001 taccggtgga tgtggaatgt gtgcgaggcc agaggccact tgtgtagcgc caagtgccca
     6061 gcggggctgc taaagcgcat gctccagact gccttgggaa aagcgcctcc cctacccggt
     6121 agaattcgaa cgctgacgtc atcaacccgc tccaaggaat cgcgggccca gtgtcactag
     6181 gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg gctgtgaggg acaggggagt
     6241 ggcgccctgc aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat
     6301 gtctttggat ttgggaatct tataagttct gtatgagacc acagatcccc gggtgcagac
     6361 aaatgtgaat tcaagagatt cacatttgtc tgcacccttt tta
//
