LOCUS       sequence-431422-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RSV-driven, lentiGuide-based destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 6538)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pRDA_512
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    453..1041
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(506..526)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(924..935)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1113..1346
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1424..1545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1461..1480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1515..1534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1585..1720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1740..1759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1741..1759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1769..1786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1769..1785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1778..1800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1927..2382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2014..2033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2224..2245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2408..2512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2513..2581,2582..3373)
                     /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(2731..2750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3544..4132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4033..4052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4286..4303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4420..4441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4456..4461,4462..4479,4480..4486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4494..4510
                     /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     4499..4521
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4518..4534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4518..4534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4553..4573
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4555..4573
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4601..4827
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4828..5008
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5055..5180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5673..5906
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6091..6135
                     /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    6369..6486
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggcaga acgtcttcac cagtaattta cgtgggaaag gtgcttgtgt cccaacaggc
       61 taggatataa tgctgaaacc gtcccccaag cgttcagggt ggggtttgct acaacttccg
      121 agtccaatgt gtccgtgttc atgatatata tgctcaaggg cgagaattgg acctagcttt
      181 cgtgttagta cgtagcatgg tgacacaagc acagtagatc ctgcccgcgt atcctatata
      241 ttaagttaat tcttatggaa tataataaca tgtggatggc cagtggtcgg ttgttacacg
      301 cctaccgcga tgctgaatga cccggactag agtggcgaaa tttatggcgt gtgacccgtt
      361 atgctccatt tcggtcagtg ggtcattgct agtagtcgat tgcattgcca ttctccgagt
      421 gatttatgga ccccagtgaa gacgcattcg aaaatcaacc tctggattac aaaatttgtg
      481 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
      541 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
      601 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
      661 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
      721 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
      781 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
      841 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
      901 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
      961 tgctgccggc tctgcggcct cttccgcgtc ctcgccttcg ccctcagacg agtcggatct
     1021 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
     1081 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     1141 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     1201 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     1261 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     1321 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca
     1381 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     1441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     1561 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     1681 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     1741 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     1801 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1861 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1921 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2341 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     2401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4201 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4261 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4321 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4381 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     4441 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     4501 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     4561 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     4621 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     4681 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     4741 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     4801 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     4861 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     4921 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     4981 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5041 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5101 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5161 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     5221 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     5281 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     5341 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     5401 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     5461 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     5521 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     5581 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     5641 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     5701 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     5761 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     5821 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     5881 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     5941 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6001 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6061 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6121 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6181 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     6241 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     6301 aggggaccct ggtacccatg catgcatgaa ttctagatct tgagacaaat ggcagtattc
     6361 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     6421 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     6481 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgaggggg
//
