LOCUS       sequence-360836-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lenti vector designed to express rtTA from a 0.5 Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  TITLE     Ploski Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7406)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti-0.5Syn-rtTA-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..259
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             260..440
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    487..612
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1105..1338
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1523..1567
                     /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    1834..1951
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             join(2516..3133,3134..3139,3140..3175,3176..3178,3179..3214,
                     3215..3217,3218..3253,3254..3262)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    3289..3877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3342..3362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3760..3771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3957..4190
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4262..4383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4299..4318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4353..4372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4423..4558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4578..4597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4579..4597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4607..4624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4607..4623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4616..4638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4765..5220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4852..4871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5062..5083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5246..5350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5351..5419,5420..6211)
                     /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(5569..5588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6382..6970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6871..6890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7124..7141
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7258..7279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7294..7299,7300..7317,7318..7324)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7332..7348
                     /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     7337..7359
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7356..7372
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7356..7372
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     join(7391..7406,1..5)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        join(7393..7406,1..5)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
       61 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
      121 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
      181 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
      241 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
      301 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
      361 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
      421 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
      481 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
      541 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
      601 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
      661 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
      721 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
      781 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
      841 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
      901 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
      961 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1021 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1081 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1141 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1201 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1261 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1321 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1381 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1441 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     1501 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     1561 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     1621 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     1681 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     1741 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     1801 attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg
     1861 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     1921 acaaaaacaa attacaaaaa ttcaaaattt tatcgataca agtatctggg aagggtaaca
     1981 atgcaagtta gcctgaggat ttaggaggag gctgaaaaac agagtaggag ccttactacg
     2041 ggtccagacc ctacggacaa gaacccccac tcccactccc caaattgcgc attccctccc
     2101 ccatcagagg gggaggggaa gaggatgcag cgcggcgcgg cgcgtgcgca ctgtcggatt
     2161 tagtaccgcg gacagagcct tcgcccccgc tgccggcgcg cgccaccacc tccccagcac
     2221 caaaggcggg ctgacgtcac tctccagccc tccccaaact cccctacctc accgccttgg
     2281 tcgcgtccgt gcagcggtga gtccagtcgg gccgcaccac aagaggtgca agataggggg
     2341 gtgcaggcgc gaccatacgc tctgcggcgg cagagcctca gcgctgcctc agtctgcagc
     2401 gggcagcaga ggagtcgcgt cgtgccagag agcgccgccg tgctcctgag ccccttgcgc
     2461 tccgcccccg cggcccaccg acccactgcc ccttggatcc actagtgccg ccaccatgtc
     2521 tagactggac aagagcaaag tcataaacgg cgctctggaa ttactcaatg gagtcggtat
     2581 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     2641 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     2701 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     2761 cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     2821 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     2881 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     2941 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     3001 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     3061 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     3121 gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca tgctcccagc
     3181 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     3241 ccttgacatg ctccccgggt aaacgcgtac cggttagtaa tgatcgacaa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gcgatggtac ctttaagacc
     3901 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     3961 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     4021 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4081 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4141 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     4201 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     4261 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4321 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4381 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     4441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4501 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4561 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     4621 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4681 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4741 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4801 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4861 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4921 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4981 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5041 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5101 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5161 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5221 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5281 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5341 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5401 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5461 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5521 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5581 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5641 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5701 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5761 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5821 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5881 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5941 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6001 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6061 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6121 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6181 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7021 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7081 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7141 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7201 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7261 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7321 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7381 cgccaagcgc gcaattaacc ctcact
//
