LOCUS       sequence-329693-        8444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector to express overexpression cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8444)
  AUTHORS   Yu M, Peng Z, Qin M, Liu Y, Wang J, Zhang C, Lin J, Dong T, Wang L, 
            Li S, Yang Y, Xu S, Guo W, Zhang X, Shi M, Peng H, Luo X, Zhang H, 
            Zhang L, Li Y, Yang XP, Sun S
  TITLE     Interferon-gamma induces tumor resistance to anti-PD-1 immunotherapy
            by promoting YAP phase separation.
  JOURNAL   Mol Cell. 2021 Feb 13. pii: S1097-2765(21)00010-1. doi: 
            10.1016/j.molcel.2021.01.010.
  PUBMED    33606996
REFERENCE   2  (bases 1 to 8444)
  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: "Mol Cell. 
            2021 Feb 13. pii: S1097-2765(21)00010-1. doi: 
            10.1016/j.molcel.2021.01.010."
COMMENT     Sequence Label: pHAGE-neo
FEATURES             Location/Qualifiers
     source          1..8444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..598
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(63..83)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(481..492)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             673..906
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1025..1043)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1111..1215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1216..1284,1285..2076)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1434..1453)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2247..2835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2736..2755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2989..3006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3246..3267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3282..3287,3288..3305,3306..3312)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3320..3336
                     /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)."
     primer_bind     3325..3347
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3344..3360
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3344..3360
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3410..3429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3621..4254
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4301..4426
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4923..5156
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5341..5385
                     /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    5678..5795
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5843..7021
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6074..7012
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6969..6989
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    7062..7629
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7079..7631
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7223..7240)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7450..7469
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7646..8440
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7700..7719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8310..8329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 atagatccta atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
       61 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
      121 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
      181 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
      241 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
      301 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
      361 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
      421 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
      481 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
      541 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
      601 gagatccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
      661 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
      721 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      781 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      841 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      901 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
      961 tatcagagag tgagaggccc gggttaatta aggaaagggc tagatcattc ttgaagacga
     1021 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1081 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1201 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1261 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1321 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1381 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1441 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1501 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1561 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1621 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1681 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1741 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1801 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1861 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1921 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1981 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2041 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2581 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2641 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2701 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2761 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2821 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2881 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2941 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3001 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3061 cattaatgca gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3121 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3181 gcaaaaagct ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3361 atgattacga atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3421 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     3481 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
     3541 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
     3601 tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac aagatatcct
     3661 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     3721 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     3781 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     3841 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     3901 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     3961 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     4021 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     4081 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4141 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     4201 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     4261 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     4321 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     4381 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     4441 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     4501 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     4561 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     4621 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     4681 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     4741 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     4801 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     4861 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     4921 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     4981 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     5041 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     5101 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     5161 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     5221 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     5281 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     5341 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     5401 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     5461 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     5521 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     5581 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     5641 tatcgccgaa ttcacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     5701 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     5761 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     5821 atccagtttg gactagtcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     5881 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     5941 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     6001 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     6061 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     6121 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
     6181 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     6241 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     6301 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     6361 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     6421 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     6481 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     6541 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     6601 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     6661 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     6721 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     6781 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     6841 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     6901 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     6961 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     7021 aagcggccgc accggtctgc agctagctcg agtctagagg atccctcccc cccccctaac
     7081 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     7141 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     7201 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     7261 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     7321 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     7381 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     7441 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     7501 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     7561 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     7621 cgatgataat atggccacac atatgatgat tgaacaagat ggattgcacg caggttctcc
     7681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7801 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     7861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     8041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     8101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     8161 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     8221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     8281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     8341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     8401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga atcg
//
