LOCUS       sequence-12973-e        6231 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 6231)
  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 6231)
  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: pLPCXU6PT7SK
FEATURES             Location/Qualifiers
     source          1..6231
                     /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             2970..3563
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3666..3688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    3816..3849
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3909..3942
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3962..3981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4062..4079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4233..4821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4313..4332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4992..5783,5784..5852))
                     /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     5615..5634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5853..5957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6025..6043
                     /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 ctgcaatcaa gctttaagga gtttatatgg
     2521 aaacccttag acgtcggcac gaggtttagc tacactcgtg ccatcgagga tgtgaggcga
     2581 tctggctgcg acatctgtca ccccattgat cgccagggtt gattcggctg atctggctgg
     2641 ctaggcgggt gtccccttcc tccctcaccg ctccatgtgc gtccctcccg aagctgcgcg
     2701 ctcggtcgaa gaggacgacc atccccgata gaggaggacc ggtcttcggt caagggtata
     2761 cgagtagctg cgctcccctg ctagaacctc caaacaagct ctcaaggtcc atttgtagga
     2821 gaacgtaggg tagtcaagct tccaagactg cagacacatc caaatgaggc gctgcatgtg
     2881 gcagtctgcc tttctttttt ctagagcggc cgcctcggcc aaacatcgat aaaataaaag
     2941 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     3001 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     3061 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     3121 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     3181 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     3241 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     3301 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     3361 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     3421 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     3481 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     3541 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     3601 gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg tgatgacggt
     3661 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3721 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     3781 atgacccagt cacgtagcga tagcggagtg tactgataac ttcgtataat gtatgctata
     3841 cgaagttatt aggtctgaag aggagtttac gtccagccaa gctagggccg cgatccggaa
     3901 cccttaatat aacttcgtat aatgtatgct atacgaagtt atcagtactg gcttaactat
     3961 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4021 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     4081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5281 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     5341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5581 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     5641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6001 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     6061 ttgctagcaa ttgctagcaa ttgctagcaa ttcataccag atcaccgaaa actgtcctcc
     6121 aaatgtgtcc ccctcacact cccaaattcg cgggcttctg cctcttagac cactctaccc
     6181 tattccccac actcaccgga gccaaagccg cggcccttcc gtttctttgc t
//
