LOCUS Exported 8063 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian Expression of p14ARF IRES zsGreen. ACCESSION . VERSION . KEYWORDS pHIV-ARF-zsGreen SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8063) 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 8063) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8063 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1684..1917 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2102..2146 /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 2444..2561 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2624..3802 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2855..3793 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3750..3770 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 4247..4833 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4414..4431) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4641..4660 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4834..5529 /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" primer_bind complement(5543..5559) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5544..5560) /label=pBluescriptKS /note="For pBluescript vector" LTR 6069..6249 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6311..6899) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6391..6410) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7070..7930) /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 7693..7712 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7931..8035) /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 gccgctgatc ttcagacctg gaggaggaga 1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact 2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg 2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 2581 gcagagatcc agtttggtta gtaccgggcc cgctctagcg tgaggctccg gtgcccgtca 2641 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg 2701 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct 2761 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt 2821 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg 2881 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca 2941 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg 3001 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg 3061 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc 3121 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa 3181 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc 3241 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat 3301 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg 3361 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag 3421 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga 3481 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc 3541 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg 3601 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga 3661 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc 3721 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt 3781 tcttccattt caggtgtcgt gagcggccgc tgaactgaat tcatggtgcg caggttcttg 3841 gtgaccctcc ggattcggcg cgcgtgcggc ccgccgcgag tgagggtttt cgtggttcac 3901 atcccgcggc tcacggggga gtgggcagcg ccaggggcgc ccgccgctgt ggccctcgtg 3961 ctgatgctac tgaggagcca gcgtctaggg cagcagccgc ttcctagaag accaggtcat 4021 gatgatgggc agcgcccgag tggcggagct gctgctgctc cacggcgcgg agcccaactg 4081 cgccgacccc gccactctca cccgacccgt gcacgacgct gcccgggagg gcttcctgga 4141 cacgctggtg gtgctgcacc gggccggggc gcggctggac gtgcgcgatg cctggggccg 4201 tctgcccgtg gacctggctg atctagagta cccgggctag gatccgcccc tctccctccc 4261 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata 4321 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg 4381 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt 4441 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag 4501 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc 4561 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga 4621 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg 4681 cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat 4741 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc 4801 tttgaaaaac acgatgataa tatggccaca accatggccc agtccaagca cggcctgacc 4861 aaggagatga ccatgaagta ccgcatggag ggctgcgtgg acggccacaa gttcgtgatc 4921 accggcgagg gcatcggcta ccccttcaag ggcaagcagg ccatcaacct gtgcgtggtg 4981 gagggcggcc ccttgccctt cgccgaggac atcttgtccg ccgccttcat gtacggcaac 5041 cgcgtgttca ccgagtaccc ccaggacatc gtcgactact tcaagaactc ctgccccgcc 5101 ggctacacct gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat ctgcaacgcc 5161 gacatcaccg tgagcgtgga ggagaactgc atgtaccacg agtccaagtt ctacggcgtg 5221 aacttccccg ccgacggccc cgtgatgaag aagatgaccg acaactggga gccctcctgc 5281 gagaagatca tccccgtgcc caagcagggc atcttgaagg gcgacgtgag catgtacctg 5341 ctgctgaagg acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa ggccaagtcc 5401 gtgccccgca agatgcccga ctggcacttc atccagcaca agctgacccg cgaggaccgc 5461 agcgacgcca agaaccagaa gtggcacctg accgagcacg ccatcgcctc cggctccgcc 5521 ttgccctaag gatcggccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg 5581 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac 5641 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt 5701 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa 5761 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact 5821 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat 5881 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga 5941 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat 6001 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg 6061 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 6121 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 6181 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 6241 tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 6301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 6361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 6421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 6481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 6541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 6601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 6661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 6721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 6781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 6841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 7021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 7141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 8041 ttccccgaaa agtgccacct gac //