LOCUS       sequence-36966-e        7586 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 7586)
  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 7586)
  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*/Neo
FEATURES             Location/Qualifiers
     source          1..7586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2557..3056
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2592..2615)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2989..3008
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3070..3873
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3133..3152)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3743..3762
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3948..4462
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4655..4671
                     /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(4679..4685,4686..4703,4704..4709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4862..4879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5792..6583,6584..6652))
                     /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     6415..6434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6653..6757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6825..6843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6881..6903)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7003..7022
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7216..7238
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7366..7385
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7585..7586,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaattc atgtcgatga gtccaaagca cacgactccg
     1441 ttctcagtgt ctgacatctt gagtcccctg gaggaaagct acaagaaagt gggcatggag
     1501 ggcggcggcc tcggggctcc gctcgcagcg tacagacagg gccaggcggc cccaccggcc
     1561 gcggccatgc agcagcacgc cgtggggcac cacggcgccg tcaccgccgc ctaccacatg
     1621 acggcggcgg gggtgcccca gctctcgcac tccgccgtgg ggggctactg caacggcaac
     1681 ctgggcaaca tgagcgagct gccgccttac caggacacca tgcggaacag cgcttcgggc
     1741 cccggatggt acggcgccaa cccagacccg cgcttccccg ccatctcccg cttcatgggc
     1801 ccggcgagcg gcatgaatat gagtggcatg ggcggcctgg gctcgctggg ggacgtgagc
     1861 aagaacatgg ccccgctgcc cagtgcgccc cgccggaagc gccgggtgct cttctcccag
     1921 gcgcaggtgt acgagctcga gcgacgtttc aagcaacaga agtacctgtc ggcgccggag
     1981 cgcgagcatc tggccagcat gattcacctg acacccacgc aggtcaagat ctggttccag
     2041 aaccaccgct acaagatgaa gcgccaggct aaggacaagg cggcgcagca acaactgcag
     2101 caggacagcg gcggcggcgg aggcggcggt ggcggtgcgg gatgcccgca gcagcagcaa
     2161 gctcagcagc agtcgccgcg ccgggtggcc gtgccggtcc tagtcaaaga cggcaaaccc
     2221 tgccaggcgg gcgcccctgc cccgggagcc gcaagcctgc aaagccacgc gcagcaacaa
     2281 gctcagcagc aggcgcaggc ggcgcaagcg gctgccgcgg ccatctcagt gggcagcggt
     2341 ggcgcggttc taggagcaca cccaggccac cagccgggca gcgcagggca gtccccggac
     2401 ctggcgcacc acgcagccag ccccgcgggg ctgcagggcc aggtctccag cctatcccat
     2461 ctgaactcct cgggctcgga ctatggcgcc atgtcctgct ccactttgct ttatggtcgg
     2521 acctggtgag ttaacctcga gagatctaat tctaccgggt aggggaggcg cttttcccaa
     2581 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     2641 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     2701 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     2761 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     2821 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     2881 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     2941 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     3001 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     3061 gcagccaata tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct
     3121 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3181 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3241 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3301 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3361 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3421 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3481 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3541 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3601 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3661 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3721 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3781 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3841 gccttctatc gccttcttga cgagttcttc tgaggggatc cgtcgacctg cagccaagct
     3901 tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3961 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     4021 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     4081 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     4141 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     4201 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     4261 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     4321 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     4381 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     4441 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     4501 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     4561 ctccaatact cctgaaatag ttcattatgg acagcgcaga agagctgggg agaattaatt
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4681 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4741 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6781 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6841 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6901 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6961 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     7021 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7081 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7141 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7201 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg
     7261 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     7321 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     7381 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     7441 gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac
     7501 tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt
     7561 atttagtctc cagaaaaagg ggggaa
//
