LOCUS       sequence-70485-e        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation Lentiviral vector with bi-cistronic expression of 
            dtomato and the gene of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  TITLE     pUltra series
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8316)
  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            "Unpublished"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: pUltra-Chili
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             236..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1835..1952
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2017..3228
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3065..3084
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3271..3972
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3404..3422)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3672..3691
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3866..3889
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3982..4038
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4060..4113
                     /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"
     misc_feature    4157..4745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4210..4230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4628..4639)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4836..5069
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5141..5262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5178..5197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5232..5251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5302..5437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5457..5476)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5458..5476)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5486..5503)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5486..5502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5495..5517)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5647..6102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5734..5753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5944..5965
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6131..6235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6236..6304,6305..7096)
                     /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(6454..6473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7267..7855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7756..7775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8009..8026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8143..8164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8179..8184,8185..8202,8203..8209)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8217..8233
                     /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     8222..8244
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8241..8257
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8241..8257
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8276..8296
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8278..8296
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acctgaaagc gaaagggaaa ccagagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     1801 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     1861 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     1921 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     1981 cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg gttttggcgc
     2041 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg
     2101 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     2161 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     2221 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     2281 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     2341 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     2401 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     2461 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     2521 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     2581 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     2641 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt
     2701 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     2761 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     2821 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     2881 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     2941 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3001 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3061 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3121 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3181 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct
     3241 gcaggtccga tccaccggtg gatcgatccg atggtgagca agggcgagga ggtcatcaaa
     3301 gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc
     3361 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     3421 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc
     3481 aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag
     3541 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag
     3601 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc
     3661 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc
     3721 ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac
     3781 ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg
     3841 cccggctact actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc
     3901 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac
     3961 gagctgtaca agggatctgg agcaacaaac ttctcactac tcaaacaagc aggtgacgtg
     4021 gaggagaatc ccgggccttc tagaggatcc ggctccggcg agggcagggg aagtctacta
     4081 acatgcgggg acgtggagga aaatcccggc ccagctagcg tgtgatcaga attcgttccg
     4141 gagtcgtcga ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4201 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4261 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4321 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4381 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4441 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4501 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     4561 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4621 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4681 gcctcttccg catcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4741 cccgcctgga attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
     4801 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctacgta actcccaacg
     4861 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     4921 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     4981 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5041 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     5101 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     5161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     5281 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5341 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5401 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     5461 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5521 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5581 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5641 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5701 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5761 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5821 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5881 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5941 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6001 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6061 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcccaggtg gcacttttcg
     6121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6421 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6541 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6781 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6841 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7081 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7381 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     7441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7981 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     8041 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     8101 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     8161 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     8221 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     8281 taaccctcac taaagggaac aaaagctgga gctgca
//
