LOCUS       sequence-406190-        7557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral eGFP reporter vector with SCP1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7557)
  AUTHORS   Taskiran II, Spanier KI, Dickmanken H, Kempynck N, Pancikova A, Eksi
            EC, Hulselmans G, Ismail JN, Theunis K, Vandepoel R, Christiaens V, 
            Mauduit D, Aerts S
  TITLE     Cell type directed design of synthetic enhancers.
  JOURNAL   Nature. 2023 Dec 12. doi: 10.1038/s41586-023-06936-2.
  PUBMED    38086419
REFERENCE   2  (bases 1 to 7557)
  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: "Nature. 
            2023 Dec 12. doi: 10.1038/s41586-023-06936-2."
COMMENT     Sequence Label: pSA351_SCP1_intron_eGFP
FEATURES             Location/Qualifiers
     source          1..7557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    193..314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(230..249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     284..303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      354..489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(509..528)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(510..528)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(538..555)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(538..554)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(547..569)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      696..1151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(783..802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     993..1014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1177..1281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1282..1350,1351..2142)
                     /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(1500..1519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2313..2901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2802..2821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3055..3072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(3225..3230,3231..3248,3249..3255)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3263..3279
                     /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     3268..3290
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3287..3303
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3287..3303
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3322..3342
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3324..3342
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3370..3596
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3597..3777
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3824..3949
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4442..4675
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4860..4904
                     /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    5171..5288
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     intron          5453..5682
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5665..5684
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      5707..5716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5713..5715,5716..5718,5719..6432)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5713..5715,5716..5718,5719..6429)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5758..5779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6019..6038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6366..6387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6448..7036
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6501..6521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6919..6930)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(7498..7557,1..121)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
       61 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      121 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
      181 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
      241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
      301 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
      361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
      421 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
      481 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
      541 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      601 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      661 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
      721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      901 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1021 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1141 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1261 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1321 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1381 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1441 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1501 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1561 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1621 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1681 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1741 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1801 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1861 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1921 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1981 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2041 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2101 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2161 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2221 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2281 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2341 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2401 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2461 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2521 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2581 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2641 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2701 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2761 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2821 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2881 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2941 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3001 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3061 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3121 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3181 aacgcaatta aggtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     3241 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     3301 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     3361 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     3421 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     3481 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     3541 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     3601 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     3661 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     3721 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     3781 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     3841 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     3901 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     3961 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     4021 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     4081 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     4141 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     4201 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     4261 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     4321 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     4381 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     4441 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     4501 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     4561 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     4621 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     4681 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     4741 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     4801 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     4861 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     4921 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     4981 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     5041 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     5101 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
     5161 atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     5221 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     5281 aaaattttat cgatcacgag actagcctcg agtcgctgag ctccctaggg tacttatata
     5341 agggggtggg ggcgcgttcg tcctcagtcg cgatcgaaca ctcgagccga gcagacgtgc
     5401 ctacggaccg gggcccgaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac
     5461 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     5521 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca
     5581 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac
     5641 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtca
     5701 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     5761 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     5821 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     5881 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     5941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     6001 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     6061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     6121 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     6181 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     6241 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     6301 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     6361 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     6421 ctgtacaagt aaagcggccg cgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     6481 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6541 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6601 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6661 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6721 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6781 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6841 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6901 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6961 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     7021 tgggccgcct ccccgcctgg aattgtacct ttaagaccaa tgacttacaa ggcagctgta
     7081 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7141 agacaagatc taaggatggg ctatgaggtg gtcgagggac catgcattct gggtgcacag
     7201 ctgaggaaaa gccccagagg atgtcccccc gcgttcatac cctaagagaa gatgcaacag
     7261 gaacaagaaa ttgcacaaca gagctctgtg gaccatgaaa aggccccttc ctccacagtg
     7321 tccctccggc gccctctact gacaaagctt gatatcgtgc ttgctgcaaa gaagggctta
     7381 gagtccagtc catcaaggag caggcactca cggggaagtt ggaagctgag aggcaagaag
     7441 aaaccagggc tctgcaaact acggccacca ttcagacgta tgctttttgc ttgtactggg
     7501 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaaccca
//
