LOCUS       sequence-138532-        7570 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 7570)
  AUTHORS   Winslow MM, Dayton TL, Verhaak RG, Kim-Kiselak C, Snyder EL, Feldser
            DM, Hubbard DD, DuPage MJ, Whittaker CA, Hoersch S, Yoon S, Crowley 
            D, Bronson RT, Chiang DY, Meyerson M, Jacks T
  TITLE     Suppression of lung adenocarcinoma progression by Nkx2-1.
  JOURNAL   Nature. 2011 May 5. 473(7345):101-4.
  PUBMED    21471965
REFERENCE   2  (bases 1 to 7570)
  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: "Nature. 
            2011 May 5. 473(7345):101-4."
COMMENT     Sequence Label: pLKO.shHmga2
FEATURES             Location/Qualifiers
     source          1..7570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /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"
     promoter        2000..2248
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2000..2020
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2171..2190
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2345..2462
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2511..3021
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2940..2958
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3043..3642
                     /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(3043..3062)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3539..3559
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3658..4246
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3711..3731)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4129..4140)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4318..4551
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4629..4750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4666..4685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4720..4739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4790..4925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4945..4964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4946..4964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4974..4991)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4974..4990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..5005)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5132..5587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5219..5238)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5429..5450
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5613..5717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5718..5786,5787..6578)
                     /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(5936..5955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6749..7337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7238..7257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7491..7508
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccagag agggcctatt tcccatgatt ccttcatatt tgcatatacg
     2041 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     2101 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     2161 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     2221 atatatcttg tggaaaggac gaggtaccgg gaaacttatc aagacgatta actcgagtta
     2281 atcgtcttga taagtttctt tttgaattct agatcttgag acaaatggca gtattcatcc
     2341 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2401 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2461 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg
     2521 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2581 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     2641 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     2701 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     2761 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     2821 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     2881 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     2941 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     3001 atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga gtacaagccc
     3061 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3121 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3181 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3241 gcggacgacg gcgccgccgt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3301 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3361 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3421 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3481 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3541 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3601 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     3661 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3721 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3781 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3841 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3901 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3961 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4021 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4081 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4141 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4201 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     4261 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4321 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4381 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4441 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4501 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     4561 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     4621 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4681 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4741 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     4801 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4861 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4921 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     5041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5101 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5161 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5221 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5281 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5341 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5401 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5461 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5521 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5581 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5641 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5701 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5761 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5821 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5881 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5941 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6001 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6061 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6121 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6181 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6241 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6301 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6361 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6421 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6481 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6541 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6601 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7201 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7261 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7321 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7381 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7441 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7501 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7561 ttcattaatg
//
