LOCUS       sequence-77808-e        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivector that expresses H2B-chFP from internal human PGK 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Huss D, Benazeraf B, Wallingford A, Filla M, Yang J, Fraser SE, 
            Lansford R
  TITLE     Transgenic quail to dynamically image amniote embryogenesis.
  JOURNAL   Development. 2015 Jul 24. pii: dev.121392.
  PUBMED    26209648
REFERENCE   2  (bases 1 to 7793)
  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: 
            "Development. 2015 Jul 24. pii: dev.121392."
COMMENT     Sequence Label: pLenti.PGK.H2B-chFP.W
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             236..416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1971..2481
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2400..2418
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2532..2909
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      2922..2931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2928..3638
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3073..3091)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3341..3360
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3647..4235
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3700..3720)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4118..4129)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4322..4555
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4627..4748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4664..4683)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4718..4737
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4767..4902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4934..4953)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4935..4953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4963..4980)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4963..4979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4972..4994)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5124..5579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5211..5230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5421..5442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5608..5712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5713..5781,5782..6573)
                     /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(5931..5950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6744..7332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7233..7252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7486..7503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7620..7641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7656..7661,7662..7679,7680..7686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7694..7710
                     /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     7699..7721
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7718..7734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7718..7734
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7753..7773
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7755..7773
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattcccac ggggttgggg
     1981 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2041 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     2101 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     2161 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     2221 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     2281 cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag cggccgggaa
     2341 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     2401 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     2461 atcaccgacc tctctcccca gggggatctc gagctcaagc ttcgaattct gcagtcgacg
     2521 gtaccgccac catgccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga
     2581 aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga
     2641 gctattccat ctatgtgtac aaggttctga agcaggtcca ccctgacacc ggcatttcgt
     2701 ccaaggccat gggcatcatg aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg
     2761 aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctccagg gagatccaga
     2821 cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta
     2881 aggccatcac caagtacacc agcgctaagg atccaccggt cgccaccatg gtgagcaagg
     2941 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     3001 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     3061 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     3121 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     3181 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     3241 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     3301 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     3361 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     3421 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     3481 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     3541 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     3601 gccactccac cggcggcatg gacgagctgt acaagtaata ctcgacaatc aacctctgga
     3661 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3721 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3781 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3841 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3901 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3961 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4021 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4081 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4141 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4201 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta
     4261 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     4321 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     4381 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4441 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4501 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     4561 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     4621 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4681 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4741 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     4801 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4861 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcgtcgaga
     4921 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     4981 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5041 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5101 cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5161 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5221 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5281 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5341 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5401 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5461 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5521 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc
     5581 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5941 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7501 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7561 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7621 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7681 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7741 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct gca
//
