LOCUS       sequence-316822-        7611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral multicistronic vector for mammalian 
            expression of three genes of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7611)
  TITLE     pUltra series
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7611)
  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            "Unpublished"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-Plus
FEATURES             Location/Qualifiers
     source          1..7611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1212
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1049..1068
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1262..1318
                     /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             1340..1393
                     /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    1437..2025
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1490..1510)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1908..1919)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2116..2349
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2421..2542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2458..2477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2512..2531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2582..2717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2737..2756)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2738..2756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2766..2783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2766..2782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2775..2797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2924..3379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3221..3242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3405..3509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3510..3578,3579..4370)
                     /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(3728..3747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4541..5129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5030..5049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5283..5300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5417..5438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5453..5458,5459..5476,5477..5483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5491..5507
                     /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     5496..5518
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5515..5531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5515..5531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5550..5570
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5552..5570
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5598..5824
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5825..6005
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6052..6177
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6670..6903
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7088..7132
                     /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    7430..7547
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
       61 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
      121 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
      181 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
      241 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
      301 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
      361 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
      421 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
      481 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
      541 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
      601 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt
      661 cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct
      721 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
      781 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
      841 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
      901 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
      961 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     1021 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     1081 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     1141 atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct
     1201 tttttgttag acgaagcttg ggctgcaggt ccgatccacc ggtgatatca tgggatctgg
     1261 agcaacaaac ttctcactac tcaaacaagc aggtgacgtg gaggagaatc ccggaccttc
     1321 tagaggatcc ggctccggcg agggcagggg aagtctacta acatgcgggg acgtggagga
     1381 aaatcccggc ccagctagcg tgtgatcaga attcgttccg gagtcgtcga ctcgacaatc
     1441 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1501 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1561 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     1621 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     1681 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     1741 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1801 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     1861 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     1921 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg catcttcgcc
     1981 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattcga
     2041 gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     2101 aagaaaaggg gggactggaa gggctacgta actcccaacg aagacaagat ctgctttttg
     2161 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     2221 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     2281 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     2341 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     2401 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     2461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2521 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2641 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2701 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     2761 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2821 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2881 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3361 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3481 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3601 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3661 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3721 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3781 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3901 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4021 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4081 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4201 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4381 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4441 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4501 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4561 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4621 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4681 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4741 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4801 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4861 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4921 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4981 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5041 ggtatcttta tagtcctgtc gggtttcgcc acctctcact tgagcgtcga tttttgtgat
     5101 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5161 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5221 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5281 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5341 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5401 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5461 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5521 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     5581 tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     5641 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     5701 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     5761 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat
     5821 aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5881 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5941 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6001 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     6061 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     6121 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     6181 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     6241 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     6301 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     6361 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     6421 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     6481 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     6541 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     6601 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     6661 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     6721 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     6781 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     6841 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     6901 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     6961 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     7021 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     7081 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     7141 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     7201 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     7261 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     7321 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     7381 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     7441 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     7501 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     7561 gggacagcag agatccagtt tggttaatta acccgtgtcg gctccagatc t
//
