LOCUS       sequence-217492-        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-CFP
FEATURES             Location/Qualifiers
     source          1..7532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     361..381
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        363..381
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        409..635
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             636..816
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    863..988
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1481..1714
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1899..1943
                     /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        2241..2489
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2241..2261
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2412..2431
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2585..2702
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2751..3261
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3180..3198
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3273..3650
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3663..3672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3669..3671,3672..3674,3675..4388)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3714..3735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3975..3994)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4322..4343
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4459..4692
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4764..4885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4801..4820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4855..4874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4925..5060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5080..5099)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5081..5099)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5109..5126)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5109..5125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5118..5140)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5267..5722
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5354..5373)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5564..5585
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5748..5852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5853..5921,5922..6713)
                     /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(6071..6090)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6884..7472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7373..7392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      361 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
      421 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      481 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      541 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
      601 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
      661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      841 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
      901 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
      961 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1021 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1081 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1141 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1201 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1261 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1321 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1381 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1441 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1501 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1561 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1621 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1681 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1741 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1801 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1861 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1921 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     1981 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2041 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2101 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2161 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc
     2221 gagcggccgc ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac
     2281 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
     2341 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     2401 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     2461 tatatatctt gtggaaagga cgaaacaccg gcaacaagat gaagagcacc aactcgagtt
     2521 ggtgctcttc atcttgttgt ttttaattct cgacctcgag acaaatggca gtattcatcc
     2581 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2641 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2701 ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag ggggttgggg
     2761 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2821 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     2881 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     2941 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     3001 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     3061 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     3121 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     3181 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     3241 atcaccgacc tctctcccca gggggatcta ccatgccaga gccagcgaag tctgctcccg
     3301 ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc
     3361 gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc
     3421 accctgacac cggcatttcg tccaaggcca tgggcatcat gaactcgttt gtgaacgaca
     3481 ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca
     3541 tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc
     3601 acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag gatccaccgg
     3661 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3721 agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc gagggcgatg
     3781 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3841 ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc taccccgacc
     3901 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgta
     3961 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4021 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4081 tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc gccgacaagc
     4141 agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4201 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4261 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4321 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4381 acaagtaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     4441 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     4501 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     4561 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     4621 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     4681 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     4741 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4861 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     4921 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4981 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5041 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     5101 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5161 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5221 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     5281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5401 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5701 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5821 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5881 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5941 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6001 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6061 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6121 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6181 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6241 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6301 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6361 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6421 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6481 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6541 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6601 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6661 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6721 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6781 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6841 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6901 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6961 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7021 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7081 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7141 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7201 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7261 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7321 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7381 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7441 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7501 tcctggcctt ttgctggcct tttgctcaca tg
//
