LOCUS       sequence-387442-        8179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid DNA for subcloning an oligonucleotide library 
            encoding RNA barcodes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8179)
  TITLE     Plasmid for generating retroviral RNA barcodes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8179)
  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            "Unpublished"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: pRV-TdTom-MCS
FEATURES             Location/Qualifiers
     source          1..8179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             670..845
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    908..1107
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1308..1724
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1654..1676
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1692..1708
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1738..2117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2361..3377
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3301..3323
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3385..3404
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3467..4897
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    4952..5540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5005..5025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5423..5434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5735..6204
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6334..6351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6505..7093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6585..6604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7253..8044,8045..8113))
                     /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     7876..7895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc
       61 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
      721 tgagtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
      781 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt
      841 tcatttgggg gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc
      901 gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg
      961 atgttatgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     1021 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     1081 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     1141 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     1201 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     1261 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     1321 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1381 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1441 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1501 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1561 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1621 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1681 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt cgatgtcgac
     1741 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     1801 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1861 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1921 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1981 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2041 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2101 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     2161 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     2221 gggcgggggg gggggggggg ggcggggcgg ggcggrgrgg kgcggcggcr gccaatcaga
     2281 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     2341 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     2401 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     2461 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     2521 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     2581 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
     2641 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     2701 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     2761 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     2821 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     2881 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     2941 kgggggtgcc gggcggggcg gggccgccyc gggccggggg gggctcgggg grggggcgcg
     3001 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     3061 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     3121 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     3181 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     3241 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     3301 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     3361 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     3421 gcaaagaatt cgctagcgga tccggccgcc tcggccaccg gtcgtcatgg tgagcaaggg
     3481 cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     3541 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3601 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     3661 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     3721 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     3781 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     3841 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     3901 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     3961 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     4021 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     4081 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     4141 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     4201 caacaacatg gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
     4261 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     4321 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     4381 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
     4441 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     4501 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
     4561 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     4621 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     4681 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
     4741 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
     4801 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
     4861 cctgttcctg tacggcatgg acgagctgta caagtaagtc tcgtttccta ggaagcttac
     4921 gcgtgcggcc gcttaattaa gcatgcaaac caatcaacct ctggattaca aaatttgtga
     4981 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5041 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5101 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5161 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5221 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5281 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5341 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5401 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5461 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5521 cctttgggcc gcctccccgc ctggaattcg agctcgagct tgttaacatc gataaaataa
     5581 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     5641 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     5701 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     5761 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     5821 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     5881 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     5941 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     6001 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     6061 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     6121 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6181 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     6241 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     6301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7201 tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8101 gttgaatact catactcttc ctttttcaat gatccggcca ttagccatat tattcattgg
     8161 ttatatagca taaatcaat
//
