LOCUS       sequence-11529-e        8039 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 8039)
  AUTHORS   Young NP, Jacks T
  TITLE     Tissue-specific p19Arf regulation dictates the response to oncogenic
            K-ras.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Jun 1. 107(22):10184-9.
  PUBMED    20479239
REFERENCE   2  (bases 1 to 8039)
  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: "Proc Natl 
            Acad Sci U S A. 2010 Jun 1. 107(22):10184-9."
COMMENT     Sequence Label: pSICO-Flpo
FEATURES             Location/Qualifiers
     source          1..8039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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"
     misc_feature    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2854..2874
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(2892..2925)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     promoter        2951..3450
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2986..3009)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3383..3402
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3464..3473
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3470..3475,3476..3496,3497..4768)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     protein_bind    complement(4777..4810)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     4837..4853
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4928..5516
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4981..5001)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5399..5410)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5519..5535)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6045..6225
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6287..6875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6367..6386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7046..7837,7838..7906))
                     /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     7669..7688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7907..8011)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg ccgccatctc
     2641 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2701 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2761 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2821 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2881 gtaaagtaat tataacttcg tatagtataa attatacgaa gttataagcc ttggtttttt
     2941 gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     3001 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     3061 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     3121 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     3181 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     3241 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     3301 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     3361 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     3421 ttctcctctt cctcatctcc gggcctttcg acctgcaatc gccgccacca tggctcctaa
     3481 gaagaagagg aaggtgatga gccagttcga catcctgtgc aagacccccc ccaaggtgct
     3541 ggtgcggcag ttcgtggaga gattcgagag gcccagcggc gagaagatcg ccagctgtgc
     3601 cgccgagctg acctacctgt gctggatgat cacccacaac ggcaccgcca tcaagagggc
     3661 caccttcatg agctacaaca ccatcatcag caacagcctg agcttcgaca tcgtgaacaa
     3721 gagcctgcag ttcaagtaca agacccagaa ggccaccatc ctggaggcca gcctgaagaa
     3781 gctgatcccc gcctgggagt tcaccatcat cccttacaac ggccagaagc accagagcga
     3841 catcaccgac atcgtgtcca gcctgcagct gcagttcgag agcagcgagg aggccgacaa
     3901 gggcaacagc cacagcaaga agatgctgaa ggccctgctg tccgagggcg agagcatctg
     3961 ggagatcacc gagaagatcc tgaacagctt cgagtacacc agcaggttca ccaagaccaa
     4021 gaccctgtac cagttcctgt tcctggccac attcatcaac tgcggcaggt tcagcgacat
     4081 caagaacgtg gaccccaaga gcttcaagct ggtgcagaac aagtacctgg gcgtgatcat
     4141 tcagtgcctg gtgaccgaga ccaagacaag cgtgtccagg cacatctact ttttcagcgc
     4201 cagaggcagg atcgaccccc tggtgtacct ggacgagttc ctgaggaaca gcgagcccgt
     4261 gctgaagaga gtgaacagga ccggcaacag cagcagcaac aagcaggagt accagctgct
     4321 gaaggacaac ctggtgcgca gctacaacaa ggccctgaag aagaacgccc cctaccccat
     4381 cttcgctatc aagaacggcc ctaagagcca catcggcagg cacctgatga ccagctttct
     4441 gagcatgaag ggcctgaccg agctgacaaa cgtggtgggc aactggagcg acaagagggc
     4501 ctccgccgtg gccaggacca cctacaccca ccagatcacc gccatccccg accactactt
     4561 cgccctggtg tccaggtact acgcctacga ccccatcagc aaggagatga tcgccctgaa
     4621 ggacgagacc aaccccatcg aggagtggca gcacatcgag cagctgaagg gcagcgccga
     4681 gggcagcatc agataccccg cctggaacgg catcatcagc caggaggtgc tggactacct
     4741 gagcagctac atcaacaggc ggatctgagc ggccgcataa cttcgtatag tataaattat
     4801 acgaagttat aagccttgtt aacgcgcggt gaccctcgag gtcgacggta tcgataagct
     4861 cgcttcacga gatcatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     4921 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4981 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5041 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5101 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5161 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5221 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5281 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5341 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5401 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5461 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     5521 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     5581 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     5641 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5701 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     5761 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     5821 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     5881 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     5941 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     6001 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     6061 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6121 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6181 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     6241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
