LOCUS       sequence-12999-e        6238 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 6238)
  AUTHORS   Hogg JR, Collins K
  TITLE     RNA-based affinity purification reveals 7SK RNPs with distinct 
            composition and regulation.
  JOURNAL   RNA. 2007 Jun . 13(6):868-80.
  PUBMED    17456562
REFERENCE   2  (bases 1 to 6238)
  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: "RNA. 2007 
            Jun . 13(6):868-80."
COMMENT     Sequence Label: pLPCXU67SKTP
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1566..2165
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1566..1585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2062..2082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        2238..2487
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2238..2258
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2410..2429
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             2977..3570
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3673..3695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    3823..3856
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3916..3949
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3969..3988
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4069..4086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4240..4828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4320..4339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4999..5790,5791..5859))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5622..5641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5860..5964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6032..6050
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc
     1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
     1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg
     2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcgc
     2221 tagcggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     2281 acaaggtctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     2341 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     2401 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2461 tatatcttgt ggaaaggacg aaacaccgtg ctgcaatcgg atgtgaggcg atctggctgc
     2521 gacatctgtc accccattga tcgccagggt tgattcggct gatctggctg gctaggcggg
     2581 tgtccccttc ctccctcacc gctccatgtg cgtccctccc gaagctgcgc gctcggtcga
     2641 agaggacgac catccccgat agaggaggac cggtcttcgg tcaagggtat acgagtagct
     2701 gcgctcccct gctagaacct ccaaacaagc tctcaaggtc catttgtagg agaacgtagg
     2761 gtagtcaagc ttccaagact gcagacacat ccaaatgagg cgctgcatgt ggcagtctgc
     2821 ctttctttgg atccggcacg aggtttagct acactcgtgc cgacgtctaa ggagtttata
     2881 tggaaaccct tatttttttt atcgattcta gagcggccgc ctcggccaaa catcgataaa
     2941 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     3001 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     3061 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     3121 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     3181 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     3241 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     3301 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     3361 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     3421 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     3481 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     3541 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     3601 gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga
     3661 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3721 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3781 cgcagccatg acccagtcac gtagcgatag cggagtgtac tgataacttc gtataatgta
     3841 tgctatacga agttattagg tctgaagagg agtttacgtc cagccaagct agggccgcga
     3901 tccggaaccc ttaatataac ttcgtataat gtatgctata cgaagttatc agtactggct
     3961 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4021 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4081 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4141 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4201 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4261 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4321 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4381 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4441 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4501 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4561 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4621 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4681 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4741 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4801 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4861 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4921 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4981 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5041 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5101 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5161 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5221 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5281 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     5341 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5401 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5461 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5521 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5581 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     5641 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5701 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5761 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5821 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5881 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5941 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6001 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6061 tcaagaattg ctagcaattg ctagcaattg ctagcaattc ataccagatc accgaaaact
     6121 gtcctccaaa tgtgtccccc tcacactccc aaattcgcgg gcttctgcct cttagaccac
     6181 tctaccctat tccccacact caccggagcc aaagccgcgg cccttccgtt tctttgct
//
