LOCUS       sequence-211417-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of p14ARF IRES H2B-mOrange2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  AUTHORS   Zeng H, Jorapur A, Shain AH, Lang UE, Torres R, Zhang Y, McNeal AS, 
            Botton T, Lin J, Donne M, Bastian IN, Yu R, North JP, Pincus L, 
            Ruben BS, Joseph NM, Yeh I, Bastian BC, Judson RL
  TITLE     Bi-allelic Loss of CDKN2A Initiates Melanoma Invasion via BRN2 
            Activation.
  JOURNAL   Cancer Cell. 2018 Jul 9;34(1):56-68.e9. doi: 
            10.1016/j.ccell.2018.05.014.
  PUBMED    29990501
REFERENCE   2  (bases 1 to 8492)
  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"; doi: 
            "10.1016/j.ccell.2018.05"; journalName: "Cancer Cell"; date: 
            "2018-07-9- 9"; volume: "34"; issue: "1"; pages: "56-68.e9"
COMMENT     Sequence Label: pHIV-ARF-mOrange2
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        622..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             838..1018
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1065..1190
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1689..1922
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2107..2151
                     /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    2449..2566
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2630..3808
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2861..3799
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3756..3776
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    4268..4841
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4291..4843
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4435..4452)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4662..4681
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4847..5224
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      5237..5246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5243..5953
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5972..5988)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5973..5989)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6498..6678
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6740..7328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6820..6839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7499..8290,8291..8359))
                     /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     8122..8141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8360..8464)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      841 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      901 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      961 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     1021 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     1081 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     1141 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     1201 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1261 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1321 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1381 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1441 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1501 acaaaagtaa gaccaccgca cagcaagcgg ccggccgcgc tgatcttcag acctggagga
     1561 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1621 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1681 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1741 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1801 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1861 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1921 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1981 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2041 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2101 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2161 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2221 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2281 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2341 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2401 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2461 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2521 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2581 ggacagcaga gatccagttt ggttagtacc gggcccgctc tagccgtgag gctccggtgc
     2641 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2701 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2761 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2821 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2881 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     2941 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     3001 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     3061 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     3121 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3181 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3241 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3301 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3361 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3421 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     3481 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     3541 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     3601 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     3661 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     3721 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     3781 tttttttctt ccatttcagg tgtcgtgagc ggccgctgaa ctgaattcat cgacgttaac
     3841 tattctagag tacccatggt gcgcaggttc ttggtgaccc tccggattcg gcgcgcgtgc
     3901 ggcccgccgc gagtgagggt tttcgtggtt cacatcccgc ggctcacggg ggagtgggca
     3961 gcgccagggg cgcccgccgc tgtggccctc gtgctgatgc tactgaggag ccagcgtcta
     4021 gggcagcagc cgcttcctag aagaccaggt catgatgatg ggcagcgccc gagtggcgga
     4081 gctgctgctg ctccacggcg cggagcccaa ctgcgccgac cccgccactc tcacccgacc
     4141 cgtgcacgac gctgcccggg agggcttcct ggacacgctg gtggtgctgc accgggccgg
     4201 ggcgcggctg gacgtgcgcg atgcctgggg ccgtctgccc gtggacctgg ctgagggcta
     4261 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4321 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4381 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4441 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     4501 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     4561 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     4621 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     4681 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     4741 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     4801 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggatgc cagagccagc
     4861 gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg cgcagaagaa
     4921 aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg tgtacaaggt
     4981 tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca tcatgaattc
     5041 gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg cgcattacaa
     5101 caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc tgctgcctgg
     5161 ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt acaccagcgc
     5221 taaggatcca ccggtcgcca ccatggtgag caagggcgag gagaataaca tggccatcat
     5281 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     5341 gatcgagggc gagggcgagg gccgccccta cgagggcttt cagaccgcta agctgaaggt
     5401 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcatt tcacctacgg
     5461 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttcaagc tgtccttccc
     5521 cgagggcttc aagtgggagc gcgtgatgaa ctacgaggac ggcggcgtgg tgaccgtgac
     5581 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     5641 cttcccctcc gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctcctccga
     5701 gcggatgtac cccgaggacg gtgccctgaa gggcaagatc aagatgaggc tgaagctgaa
     5761 ggacggcggc cactacacct ccgaggtcaa gaccacctac aaggccaaga agcccgtgca
     5821 gctgcccggc gcctacatcg tcgacatcaa gttggacatc acctcccaca acgaggacta
     5881 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     5941 gctgtacaag taagcggccg cgactctaga tcgataccgt cgacctcgat cgagacctag
     6001 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     6061 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     6121 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6181 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     6241 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     6301 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     6361 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     6421 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     6481 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6721 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6781 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6841 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6901 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6961 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7021 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7081 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7141 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7201 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7261 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7321 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7381 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7441 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7501 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7561 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7621 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7681 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7741 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7801 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7861 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7921 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7981 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8041 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8101 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8161 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8221 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8281 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8341 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8401 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8461 cgcgcacatt tccccgaaaa gtgccacctg ac
//
