LOCUS       sequence-381527-        7539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  AUTHORS   Beronja S, Livshits G, Williams S, Fuchs E
  TITLE     Rapid functional dissection of genetic networks via tissue-specific 
            transduction and RNAi in mouse embryos.
  JOURNAL   Nat Med. 2010 Jul . 16(7):821-7.
  PUBMED    20526348
REFERENCE   2  (bases 1 to 7539)
  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: "Nat Med. 
            2010 Jul . 16(7):821-7."
COMMENT     Sequence Label: LV-RFP
FEATURES             Location/Qualifiers
     source          1..7539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..511
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     430..448
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             523..900
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      913..922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             919..1596
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             919..1596
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1049..1067)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1317..1336
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1511..1534
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     LTR             1716..1949
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2021..2142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2058..2077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2112..2131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2182..2317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2337..2356)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2338..2356)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2366..2383)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2366..2382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2375..2397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2524..2979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2821..2842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3005..3109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3110..3178,3179..3970)
                     /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(3328..3347)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4141..4729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4630..4649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4883..4900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5017..5038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5053..5058,5059..5076,5077..5083)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5091..5107
                     /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     5096..5118
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5115..5131
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5115..5131
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5150..5170
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5152..5170
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5198..5424
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5425..5605
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5652..5777
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6270..6503
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6688..6732
                     /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"
     promoter        7030..7278
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7030..7050
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7201..7220
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    7374..7491
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
       61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
      121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
      181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
      241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
      301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
      361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
      421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
      481 cgttgaccga atcaccgacc tctctcccca gggggatcta ccatgccaga gccagcgaag
      541 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
      601 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
      661 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaactcgttt
      721 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
      781 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
      841 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
      901 gatccaccgg tcgccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc
      961 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1021 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     1081 cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc ctacgtgaag
     1141 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1201 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1261 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1321 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta ccccgaggac
     1381 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac
     1441 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag
     1501 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1561 gagcgcgccg agggccgcca ctccaccggc gcctaactcg agactagtcg acgtgaggat
     1621 cctctagagt ccctaggatg catcggtacc tttaagacca atgacttaca aggcagctgt
     1681 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     1741 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     1801 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     1861 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     1921 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     1981 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     2041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2101 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     2161 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2281 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     2341 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2401 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2461 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2521 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2581 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2701 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2821 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2881 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2941 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     3001 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3061 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3121 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3181 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3241 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3301 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3361 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3421 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3481 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3541 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3601 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3661 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3721 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3781 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3841 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3901 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3961 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4801 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4861 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4921 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4981 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5041 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5101 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     5161 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     5221 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     5281 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     5341 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     5401 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     5461 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5521 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5581 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     5641 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     5701 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     5761 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     5821 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     5881 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     5941 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     6001 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     6061 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     6121 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     6181 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     6241 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     6301 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     6361 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     6421 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     6481 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     6541 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     6601 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     6661 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     6721 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     6781 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     6841 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     6901 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     6961 attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc
     7021 cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     7081 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
     7141 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     7201 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     7261 tggaaaggac gaaacaccgg caacaagatg aagagcacca actcgagttg gtgctcttca
     7321 tcttgttgtt tttaattctc gacctcgaga caaatggcag tattcatcca caattttaaa
     7381 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     7441 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     7501 tacagggaca gcagagatcc actttggccg cggctcgag
//
