LOCUS       sequence-163457-        6698 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 6698)
  TITLE     Viral vectors.
REFERENCE   2  (bases 1 to 6698)
  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"
COMMENT     Sequence Label: pMKO.1 puro PTEN shRNA
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        125..454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      305..440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             464..1063
                     /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(464..483)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     960..980
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1259..1684
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(1787..1809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1950..2279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2130..2265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2301..2320
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2692..3280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2772..2791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3451..4242,4243..4311))
                     /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     4074..4093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4312..4416)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4694..4712
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4812..5191
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5192..5395
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5346..5366
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5396..5571
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5634..5833
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6034..6450
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6380..6402
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6418..6434
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        join(6486..6698,1..36)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6486..6506
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6657..6676
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ttggctttat atatcttgtg gaaaggacga aacaccggac ttgaaggcgt atacaggact
       61 cgagtcctgt atacgccttc aagtcttttt gaattcgcca gcacagtggt cgaccctgtg
      121 gaatgtgtgt cagttagggt gtggaaagtc cccaggcttc ccagcaggca gaagtatgca
      181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      421 aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgaccg agtacaagcc
      481 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
      541 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
      601 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
      661 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
      721 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
      781 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
      841 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
      901 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
      961 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     1021 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     1081 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg attagtccaa tttgttaaag
     1141 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     1201 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     1261 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     1321 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     1381 ggtcgaccct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     1441 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     1501 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     1561 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     1621 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     1681 catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg
     1741 ggaggtaagc tggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1801 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1861 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     1921 atagcggagt gtagatccgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     1981 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2281 agcttactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     2341 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     2401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatggcttc
     2521 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3121 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3541 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3601 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3661 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3721 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3781 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3841 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3901 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3961 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4021 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4081 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4141 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4201 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4261 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4441 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4501 ttttttaacc aataggccga aatcggcaaa atcccttaat tattgaagca tttatcaggg
     4561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4621 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4681 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tagcttggcc
     4741 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
     4801 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4861 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4921 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4981 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5041 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5101 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5161 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     5221 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     5281 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5341 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata
     5401 aaagagccca caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt
     5461 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct
     5521 tgggagggtc tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc
     5581 gtccgggatt tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     5641 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg
     5701 cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     5761 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
     5821 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
     5881 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
     5941 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
     6001 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     6061 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     6121 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     6181 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     6241 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     6301 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     6361 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     6421 tatccagccc tcactccttc tctaggcgcc ggaattgaag atctgtcgac aaggtcgggc
     6481 aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     6541 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6601 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6661 atcatatgct taccgtaact tgaaagtatt tcgatttc
//
