LOCUS       sequence-424829-        7711 bp    DNA     circular SYN 15-APR-2025
DEFINITION  6xGCN4pep transgene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  TITLE     Cheeseman lab plasmids Nov 2023
REFERENCE   2  (bases 1 to 7711)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGS399_myc-6xGCN4-T2A-BSD Lenti
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    94..115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(130..135,136..153,154..160)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    168..184
                     /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     173..195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     192..208
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     192..208
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     227..247
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        229..247
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        275..501
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             502..682
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    729..854
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1351..1584
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1769..1813
                     /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     1991..2011
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2212..2329
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2401..2612
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2641..2670
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2713..2733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2746..2790
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2827..2892
                     /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             2908..2973
                     /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             2989..3054
                     /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             3070..3135
                     /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             3151..3216
                     /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             3232..3297
                     /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             3334..3387
                     /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             3394..3792
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3838..4426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3891..3911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4309..4320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4498..4731
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4809..4930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4846..4865)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4900..4919
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4970..5105
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5125..5144)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5126..5144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5154..5171)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5154..5170)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5163..5185)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5312..5767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5399..5418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5609..5630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5793..5897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5898..5966,5967..6758)
                     /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(6116..6135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6929..7517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7418..7437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7671..7688
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
       61 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      121 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      181 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
      241 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
      301 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
      361 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
      421 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
      481 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
      541 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
      601 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
      661 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
      721 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
      781 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
      841 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
      901 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
      961 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     1021 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     1081 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     1141 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc
     1201 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     1261 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     1321 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     1381 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     1441 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     1501 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     1561 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     1621 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     1681 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     1741 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     1801 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     1861 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     1921 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     1981 gggacccaga gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     2041 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     2101 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     2161 ggactatcat aaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     2221 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2281 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2341 cagggacagc agagatccac tttggcgccg gctcgtcgag tggctccggt gcccgtcagt
     2401 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2461 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2521 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2581 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgatatcatg
     2641 gagcagaagc tgatcagcga ggaggacctg ggaggggatc ggtggagctc caccggtggc
     2701 ggccgctcta gagagaatct ttattttcag ggcgccgcca aattcaaaga aaccgctgct
     2761 gctaaattcg aacgccagca catggacagc ggaggtggag gtactagtct cgagggttct
     2821 ggaggattgt tgcccaaaaa ctaccacttg gagaacgagg tcgcaaggtt gaaaaaattg
     2881 gtcggtgagc gtggcggctc aggcggcctt cttccaaaaa attatcatct tgaaaatgaa
     2941 gttgctagac ttaaaaaact tgttggagaa agaggcggga gcggcggcct gctgcccaag
     3001 aactaccacc tggagaacga ggtcgcccgc ctgaagaagc tggtcggcga gcgcggtggt
     3061 tctggaggtt tgctcccaaa gaactaccac ttggaaaacg aagtcgcaag attgaagaag
     3121 ttggtcggtg aaagaggagg aagcggagga ttattaccta aaaattatca tttggaaaat
     3181 gaagttgcta gattgaagaa gttggttgga gagagagggg ggtctggagg actactaccg
     3241 aaaaactacc acctagagaa cgaggtggcc cgactaaaaa agctagtggg cgagcgcggc
     3301 ggcagtggcg gcctcctccc caagaatgaa ttcgagggca gaggaagtct tctaacatgc
     3361 ggtgacgtgg aggagaatcc cggccctatc gatatggcca agcctttgtc tcaagaagaa
     3421 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     3481 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     3541 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     3601 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     3661 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     3721 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     3781 tgggagggct aaggtaccac tagtcccggg atgcatagcg ctacgcgtta agtcgacaat
     3841 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3901 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3961 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4021 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4081 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4141 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4201 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4261 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4321 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4381 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     4441 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4501 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4561 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4621 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4681 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     4741 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     4801 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4861 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4921 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     4981 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5041 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5101 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     5161 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     5221 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5281 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5341 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5401 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5461 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5521 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5581 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5641 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5701 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5761 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7501 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7561 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7621 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7681 tgagcgagga agcggaagag cgcccaatac g
//
