LOCUS       sequence-316820-        8706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral bicistronic vector for mammalian 
            expression of H2B-mTagBFP2 and one gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8706)
  TITLE     pUltra series
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8706)
  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            "Unpublished"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-H2B-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..8706
                     /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             1325..1702
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             2435..2488
                     /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    2532..3120
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2585..2605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3003..3014)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3211..3444
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3516..3637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3553..3572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3607..3626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3832..3851)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3833..3851)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3861..3878)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3861..3877)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3870..3892)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4019..4474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4106..4125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4316..4337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4500..4604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4605..4673,4674..5465)
                     /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(4823..4842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5636..6224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6125..6144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6378..6395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6512..6533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6548..6553,6554..6571,6572..6578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6586..6602
                     /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     6591..6613
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6610..6626
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6610..6626
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6645..6665
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6647..6665
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6693..6919
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6920..7100
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7147..7272
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7765..7998
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8183..8227
                     /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    8525..8642
                     /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 tagaatgcca gagccagcga agtctgctcc cgccccgaaa aagggctcca agaaggcggt
     1381 gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg agagctattc
     1441 catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt cgtccaaggc
     1501 catgggcatc atgaactcgt ttgtgaacga cattttcgag cgcatcgcag gtgaggcttc
     1561 ccgcctggcg cattacaaca agcgctcgac catcacctcc agggagatcc agacggccgt
     1621 gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta ctaaggccat
     1681 caccaagtac accagcgcta aggaccctcc agtcgccacc atgagcgagt tgatcaagga
     1741 aaacatgcat atgaagctct acatggaagg caccgtggac aaccaccatt tcaagtgtac
     1801 ctccgaggga gagggaaagc cttacgaggg aactcagacc atgcgcatca aggtcgtgga
     1861 aggcggtcca ctgcccttcg ctttcgacat tctcgccacc tcattcctct acggctccaa
     1921 gactttcatc aatcataccc agggtattcc cgatttcttc aagcaatcgt tcccggaagg
     1981 gtttacctgg gaacgcgtga ccacttacga ggacggaggg gtcctgaccg caactcagga
     2041 cactagcctg caggacggtt gcctgatcta taacgtgaag attcggggag tgaacttcac
     2101 ctccaacgga cctgtcatgc aaaagaaaac cctgggctgg gaagccttta ccgagactct
     2161 gtatccggcc gacggcggac ttgaggggag aaatgatatg gcgctgaaac tggtcggcgg
     2221 ctcccacctg atcgccaacg ccaagacgac ctaccggtcg aagaagcccg ccaagaacct
     2281 gaagatgccg ggagtgtact acgtggacta caggctggag cggatcaagg aagccaacaa
     2341 cgaaacatac gtggaacagc acgaagtggc cgtggcgcgc tactgcgatc tccctagcaa
     2401 actggggcac aaactcaatg gatccggctc cggcgagggc aggggaagtc tactaacatg
     2461 cggggacgtg gaggaaaatc ccggcccagc tagcgtgtga tcagaattcg ttccggagtc
     2521 gtcgactcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     2941 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3061 ttccgcatct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3121 ctggaattaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     3181 ttagccactt tttaaaagaa aaggggggac tggaagggct acgtaactcc caacgaagac
     3241 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3301 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3361 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3421 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     3481 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     3541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     3841 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3901 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3961 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4021 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4081 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4141 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4201 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4261 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4321 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4381 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4441 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4681 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4741 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4801 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4861 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4921 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4981 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5041 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5101 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5161 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5221 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5281 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5341 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5401 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5461 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5581 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tcacttgagc
     6181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6301 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6361 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6421 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6481 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6541 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6601 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     6661 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     6721 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     6781 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     6841 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     6901 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     6961 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7021 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7081 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     7141 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     7201 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     7261 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     7321 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     7381 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     7441 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     7501 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     7561 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     7621 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     7681 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     7741 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     7801 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     7861 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     7921 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     7981 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     8041 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     8101 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     8161 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8221 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8281 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8341 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     8401 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     8461 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     8521 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     8581 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     8641 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaacccg tgtcggctcc
     8701 agatct
//
