LOCUS       sequence-9928-ex        8264 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 8264)
  TITLE     Garbers lab plasmids
REFERENCE   2  (bases 1 to 8264)
  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"
COMMENT     Sequence Label: pLLU2G
FEATURES             Location/Qualifiers
     source          1..8264
                     /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"
     promoter        2622..2935
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2854..2874
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2952..2968
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3010..3043)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3086..4295
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4132..4151
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      4319..4328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4325..4327,4328..4330,4331..5044)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4325..4327,4328..4330,4331..5041)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4370..4391)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4631..4650)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4978..4999
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5064..5097)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5153..5741
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5206..5226)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5624..5635)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5744..5760)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6270..6450
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6512..7100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6592..6611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7271..8062,8063..8131))
                     /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     7894..7913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8132..8236)
                     /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 tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgttaacgc
     2941 gcggtgaccc tcgaggtcga cggtatcgat aagctcgctt cacgagattc cagcaggtcg
     3001 agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttccaagct
     3061 taagcggccc gcgggcccgg gatctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     3121 gcccccctcc tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc
     3181 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
     3241 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
     3301 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
     3361 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
     3421 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
     3481 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
     3541 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
     3601 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
     3661 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
     3721 aagaacccaa ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
     3781 gctggggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     3841 gtcgtctgtt gcgggggcgg cagttatgcg gtgccgttgg gcagtgcacc cgtacctttg
     3901 ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt
     3961 ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg
     4021 gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag
     4081 tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc
     4141 ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt
     4201 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg
     4261 ctaaattctg gccgtttttg gcttttttgt tagacgaagc ttgctagcgc taccggtcgc
     4321 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4381 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4441 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4501 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     4561 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4621 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4681 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4741 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     4801 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     4861 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4921 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     4981 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5041 gtaggaattc gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca
     5101 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     5161 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5221 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5281 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5341 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5401 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5461 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5521 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5581 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5641 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5701 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     5761 atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     5821 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     5881 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5941 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     6001 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     6061 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     6121 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     6181 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     6241 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     6301 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6361 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6421 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
     6481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgac
//
