LOCUS       sequence-133514-        8562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFUGW-CMVp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8562)
  AUTHORS   Wong AS, Choi GC, Cheng AA, Purcell O, Lu TK
  TITLE     Massively parallel high-order combinatorial genetics in human cells.
  JOURNAL   Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub 
            2015 Aug 17.
  PUBMED    26280411
REFERENCE   2  (bases 1 to 8562)
  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: "10.1038/nbt.3326"; 
            journalName: "Nat Biotechnol"; date: "2015-09"; volume: "33"; issue:
            "9"; pages: "952-61"
COMMENT     Sequence Label: pAWp11
FEATURES             Location/Qualifiers
     source          1..8562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..630
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(95..115)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(513..524)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(633..649)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(634..650)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1155..1335
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1361..1378)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1367..1591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1637..2065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1724..1743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1934..1955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2074..2094)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2079..2408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2259..2394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2321..2340
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2456..2503
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2522..2896
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3026..3147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3063..3082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3117..3136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3196..3212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3196..3212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3209..3231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3220..3236
                     /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(3244..3250,3251..3268,3269..3274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3289..3310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3427..3444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3598..4186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3678..3697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4357..5148,5149..5217))
                     /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     4980..4999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5218..5322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5397..5416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5588..5967
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5969..6167
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6185..6365
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6412..6537
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7030..7263
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7448..7492
                     /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    7790..7907
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        7971..8350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8352..8550
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gatccgcgaa aagtccggaa ttcgatatca agcttatcga taatcaacct ctggattaca
       61 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
      121 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
      181 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
      241 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
      301 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
      361 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
      421 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
      481 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
      541 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
      601 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa
      661 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
      721 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
      781 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
      841 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
      901 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
      961 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     1021 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     1081 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     1141 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     1201 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     1261 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     1321 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     1381 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1441 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1501 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     1561 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     1621 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1681 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1741 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1801 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1861 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1921 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1981 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2041 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2101 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2161 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2221 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2281 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2341 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2401 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     2461 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2521 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     2581 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     2641 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     2701 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     2761 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     2821 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     2881 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     2941 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3001 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3061 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3121 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3181 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3241 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3301 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3361 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3421 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3481 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3541 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3601 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3661 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3721 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3781 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3841 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3901 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3961 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4021 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4081 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4141 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4201 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4261 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4321 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4381 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4441 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4501 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4561 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4621 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4681 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4741 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4801 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4861 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4921 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4981 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5041 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5101 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5161 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5221 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5281 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5341 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     5401 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     5461 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     5521 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     5581 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     5641 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5701 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5761 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     5821 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5881 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5941 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     6001 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6061 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6121 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg
     6181 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6241 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6301 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6361 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     6421 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     6481 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     6541 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     6601 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     6661 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     6721 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     6781 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     6841 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     6901 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     6961 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     7021 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     7081 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     7141 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     7201 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     7261 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     7321 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     7381 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     7441 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     7501 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     7561 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     7621 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     7681 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     7741 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     7801 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     7861 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     7921 gggacagcag agatccagtt tggttaatta acgggccaga tatacgcgtt gacattgatt
     7981 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     8041 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     8101 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     8161 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     8221 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     8281 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     8341 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     8401 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     8461 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     8521 gcgtgtacgg tgggaggtct atataagcag cgcctgcagg cg
//
