LOCUS       sequence-395416-        6922 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 6922)
  AUTHORS   Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz 
            L, Nusse R, Weissman IL
  TITLE     A role for Wnt signalling in self-renewal of haematopoietic stem 
            cells.
  JOURNAL   Nature. 2003 May 22. 423(6938):409-14.
  PUBMED    12717450
REFERENCE   2  (bases 1 to 6922)
  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. 
            2003 May 22. 423(6938):409-14."
COMMENT     Sequence Label: Lentiviral-FOP-dGFP-reporter
FEATURES             Location/Qualifiers
     source          1..6922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2555..2574
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2555..2573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2594..2729
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2769..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2780..2799)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2834..2853
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2962..3195
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3282..3870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3388..3399
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3797..3817
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3891..4013,4014..4019,4020..4730,4731..4733,
                     4734..4736))
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(4062..4083)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4411..4430
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4670..4691
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4733..4742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4758..4767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5101..5145)
                     /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    5330..5563
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6056..6181
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6228..6408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6409..6635)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(6663..6683)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6663..6681)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6702..6718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6702..6718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6715..6737)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6726..6742
                     /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)."
     promoter        complement(join(6750..6756,6757..6774,6775..6780))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6795..6816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     2581 gggtacgtcc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     2641 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2701 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggat agctagagcc
     2761 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2821 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2881 taaacaagtt cctctcactc tctgatattc atttctttgc aagttataaa tactgaataa
     2941 taagatgaca tgaactacta ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     3001 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     3061 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     3121 tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt
     3181 gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct
     3241 tgtaagtcat tggtcttaaa ggtaccgagc tcgaattcca ggcggggagg cggcccaaag
     3301 ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag
     3361 caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt
     3421 cccgcgcaga atccaggtgg caacacaggc gagcagccat ggaaaggacg tcagcttccc
     3481 cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
     3541 gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag
     3601 gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca
     3661 caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga
     3721 tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat
     3781 taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct
     3841 ttcacaaatt ttgtaatcca gaggttgatt gtcgacctgc aggcggccgc ctacacattg
     3901 atcctagcag aagcacaggc tgcagggtga cggtccatcc cgctctcctg ggcacaagac
     3961 atgggcagcg tgccatcatc ctgctcctcc acctccggcg ggaagccatg gctaagcttc
     4021 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     4081 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     4141 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     4201 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     4261 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     4321 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     4381 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     4441 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     4501 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     4561 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     4621 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     4681 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
     4741 gcgaccggtg gatcatccca tggtggcttt accaacagta ccggaatgcc aagcttactt
     4801 agatcgcaga tctcgagtac cggatccatt atataccctc tagacctttg gccttacccc
     4861 ctttggcctt accccctttg gccctgcagg catgcaagct tcagctgctc gagcccgggc
     4921 tatgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     4981 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     5041 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     5101 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     5161 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     5221 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     5281 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     5341 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     5401 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     5461 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     5521 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     5581 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     5641 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg
     5701 tctgaagatc agcggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta
     5761 tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg
     5821 ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta
     5881 gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga
     5941 atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc
     6001 ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc
     6061 tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca
     6121 gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga
     6181 gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt
     6241 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
     6301 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
     6361 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccgt ttatgtatcg
     6421 agctaggcac ttaaatacaa tatctctgca atgcggcaat tcagtggttc gtccaatcca
     6481 tgtcagaccc gtctgttgcc ttcctaataa ggcacgatcg taccacctta cttccaccaa
     6541 tcggcatgca cggtgctttt tctctccttg taaggcatgt tgctaactca tcgttaccat
     6601 gttgcaagac tacaagagta ttgcataaga ctacattaag cttgcagctc cagcttttgt
     6661 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6901 ggcggtttgc gtattgggcg ct
//
