LOCUS       sequence-305765-        7388 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 7388)
  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 7388)
  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: MSCV-Nkx2-1/Puro
FEATURES             Location/Qualifiers
     source          1..7388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2564..3063
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2599..2622)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2996..3015
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3084..3683
                     /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(3084..3103)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3580..3600
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3783..4297
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4459..4475
                     /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)."
     promoter        complement(join(4483..4489,4490..4507,4508..4513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4666..4683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4837..5425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4917..4936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5596..6387,6388..6456))
                     /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     6219..6238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6457..6561)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6629..6647
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6685..6707)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6807..6826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7020..7042
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7170..7189
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctctc gaggttaacc caccatgtcg
     1441 atgagtccaa agcacacgac tccgttctca gtgtctgaca tcttgagtcc cctggaggaa
     1501 agctacaaga aagtgggcat ggagggcggc ggcctcgggg ctccgctcgc agcgtacaga
     1561 cagggccagg cggccccacc ggccgcggcc atgcagcagc acgccgtggg gcaccacggc
     1621 gccgtcaccg ccgcctacca catgacggcg gcgggggtgc cccagctctc gcactccgcc
     1681 gtggggggct actgcaacgg caacctgggc aacatgagcg agctgccgcc ttaccaggac
     1741 accatgcgga acagcgcttc gggccccgga tggtacggcg ccaacccaga cccgcgcttc
     1801 cccgccatct cccgcttcat gggcccggcg agcggcatga atatgagtgg catgggcggc
     1861 ctgggctcgc tgggggacgt gagcaagaac atggccccgc tgcccagtgc gccccgccgg
     1921 aagcgccggg tgctcttctc ccaggcgcag gtgtacgagc tcgagcgacg tttcaagcaa
     1981 cagaagtacc tgtcggcgcc ggagcgcgag catctggcca gcatgattca cctgacaccc
     2041 acgcaggtca agatctggtt ccagaaccac cgctacaaga tgaagcgcca ggctaaggac
     2101 aaggcggcgc agcaacaact gcagcaggac agcggcggcg gcggaggcgg cggtggcggt
     2161 gcgggatgcc cgcagcagca gcaagctcag cagcagtcgc cgcgccgggt ggccgtgccg
     2221 gtcctagtca aagacggcaa accctgccag gcgggcgccc ctgccccggg agccgcaagc
     2281 ctgcaaagcc acgcgcagca acaagctcag cagcaggcgc aggcggcgca agcggctgcc
     2341 gcggccatct cagtgggcag cggtggcgcg gttctaggag cacacccagg ccaccagccg
     2401 ggcagcgcag ggcagtcccc ggacctggcg caccacgcag ccagccccgc ggggctgcag
     2461 ggccaggtct ccagcctatc ccatctgaac tcctcgggct cggactatgg cgccatgtct
     2521 tgttctacct tgctttatgg tcggacctgg tgagaattct accgggtagg ggaggcgctt
     2581 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     2641 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     2701 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     2761 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     2821 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     2881 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     2941 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3001 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3061 tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     3121 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     3181 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     3241 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     3301 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     3361 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     3421 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     3481 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     3541 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     3601 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     3661 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     3721 gcgcacgacc ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3781 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3841 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3901 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3961 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     4021 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     4081 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     4141 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     4201 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4261 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     4321 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     4381 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     4441 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6601 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6661 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6721 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6781 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6841 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6901 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6961 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7021 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     7081 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7141 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7201 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     7261 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     7321 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     7381 aagggggg
//
