LOCUS       sequence-424828-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4xGCN4pep transgene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7543)
  TITLE     Cheeseman lab plasmids Nov 2023
REFERENCE   2  (bases 1 to 7543)
  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: pGS398_myc-4xGCN4-T2A-BSD Lenti
FEATURES             Location/Qualifiers
     source          1..7543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    104..125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(140..145,146..163,164..170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    178..194
                     /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     183..205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     202..218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     237..257
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        239..257
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        285..511
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             512..692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    739..864
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1361..1594
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1779..1823
                     /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"
     primer_bind     2001..2021
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2222..2339
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2411..2622
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2651..2680
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2723..2743
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2756..2800
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2837..2902
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             2918..2983
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             2999..3064
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             3080..3145
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             3176..3229
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3236..3634
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3680..4268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3733..3753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4151..4162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4340..4573
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4651..4772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4688..4707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4742..4761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4812..4947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4967..4986)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4968..4986)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4996..5013)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4996..5012)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5005..5027)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5154..5609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5241..5260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5451..5472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5635..5739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5740..5808,5809..6600)
                     /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     complement(5958..5977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6771..7359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7260..7279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7513..7530
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
       61 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      121 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      181 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
      241 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
      301 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
      361 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
      421 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
      481 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
      541 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      601 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      661 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
      721 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
      781 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
      841 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
      901 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
      961 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     1021 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     1081 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     1141 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     1201 gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1261 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1321 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1381 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1441 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1501 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1561 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1621 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1681 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1741 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1801 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     1861 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     1921 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     1981 caaccccgag gggacccaga gagggcctat ttcccatgat tccttcatat ttgcatatac
     2041 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
     2101 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     2161 gttttaaaat ggactatcat aaattctaga tcttgagaca aatggcagta ttcatccaca
     2221 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     2281 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     2341 gggtttatta cagggacagc agagatccac tttggcgccg gctcgtcgag tggctccggt
     2401 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2461 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2521 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2581 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc
     2641 cgatatcatg gagcagaagc tgatcagcga ggaggacctg ggaggggatc ggtggagctc
     2701 caccggtggc ggccgctcta gagagaatct ttattttcag ggcgccgcca aattcaaaga
     2761 aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag gtactagtct
     2821 cgagggttct ggaggattgt tgcccaaaaa ctaccacttg gagaacgagg tcgcaaggtt
     2881 gaaaaaattg gtcggtgagc gtggcggctc aggcggcctt cttccaaaaa attatcatct
     2941 tgaaaatgaa gttgctagac ttaaaaaact tgttggagaa agaggcggga gcggcggcct
     3001 gctgcccaag aactaccacc tggagaacga ggtcgcccgc ctgaagaagc tggtcggcga
     3061 gcgcggtggt tctggaggtt tgctcccaaa gaactaccac ttggaaaacg aagtcgcaag
     3121 attgaagaag ttggtcggtg aaagaggagg aagcggagga ttattacctg aattcgaggg
     3181 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta tcgatatggc
     3241 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     3301 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     3361 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     3421 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     3481 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     3541 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     3601 attgctgccc tctggttatg tgtgggaggg ctaaggtacc actagtcccg ggatgcatag
     3661 cgctacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3721 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3781 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3841 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3901 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3961 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4021 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     4081 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4141 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4201 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4261 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     4321 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     4381 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4441 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4501 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4561 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
     4621 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     4681 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4741 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     4801 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4861 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4921 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     4981 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5041 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5101 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     5161 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5221 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5281 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5341 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5401 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5461 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5521 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5581 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5761 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5821 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5881 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5941 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6001 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6061 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6121 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6181 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6241 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6301 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6361 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6421 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6481 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6541 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6601 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6661 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6721 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6781 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6841 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6901 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6961 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7021 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7081 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7141 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7201 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7261 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7321 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7381 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7441 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7501 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agc
//
