LOCUS       sequence-261202-        7532 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 7532)
  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 7532)
  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-YFP
FEATURES             Location/Qualifiers
     source          1..7532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             519..896
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      909..918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(915..917,918..920,921..1634)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(960..981)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1221..1240)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1568..1589
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1705..1938
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2010..2131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2047..2066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2101..2120
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2171..2306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2326..2345)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2327..2345)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2355..2372)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2355..2371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2364..2386)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2513..2968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2600..2619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2810..2831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2994..3098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3099..3167,3168..3959)
                     /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(3317..3336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4130..4718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4619..4638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4872..4889
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5006..5027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5042..5047,5048..5065,5066..5072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5080..5096
                     /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     5085..5107
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5104..5120
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5104..5120
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5139..5159
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5141..5159
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5187..5413
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5414..5594
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5641..5766
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6259..6492
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6677..6721
                     /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        7019..7267
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7019..7039
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7190..7209
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    7363..7480
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(7529..7532,1..507)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatctaccat gccagagcca gcgaagtctg
      541 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
      601 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
      661 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaac tcgtttgtga
      721 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct
      781 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
      841 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaaggatc
      901 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      961 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1021 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1081 tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc
     1141 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1201 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1261 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1321 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1381 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1441 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1501 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     1561 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1621 agctgtacaa gtaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     1681 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     1741 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     1801 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     1861 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     1921 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     1981 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     2041 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2101 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     2161 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2221 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2281 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     2341 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2401 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2461 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2641 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2881 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2941 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     3001 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3061 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3121 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3181 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3241 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3301 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3361 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3421 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3481 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3541 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3601 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3661 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3721 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3781 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3841 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3901 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3961 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4021 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4081 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4141 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4201 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4261 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4321 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4381 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4441 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4501 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4561 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4621 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4681 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4741 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4801 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4861 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4921 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4981 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5041 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5101 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     5161 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
     5221 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
     5281 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
     5341 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
     5401 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5461 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5521 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5581 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     5641 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     5701 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     5761 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     5821 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     5881 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     5941 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     6001 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     6061 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     6121 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     6181 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     6241 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     6301 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     6361 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     6421 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     6481 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     6541 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     6601 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     6661 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     6721 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     6781 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     6841 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     6901 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     6961 gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc ccttcaccga
     7021 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     7081 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     7141 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     7201 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     7261 aaacaccggc aacaagatga agagcaccaa ctcgagttgg tgctcttcat cttgttgttt
     7321 ttaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     7381 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     7441 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     7501 cagagatcca ctttggccgc ggctcgaggg gg
//
