LOCUS       sequence-212625-        7514 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 7514)
  AUTHORS   Cui Y, Golob J, Kelleher E, Ye Z, Pardoll D, Cheng L
  TITLE     Targeting transgene expression to antigen-presenting cells derived 
            from lentivirus-transduced engrafting human hematopoietic 
            stem/progenitor cells.
  JOURNAL   Blood. 2002 Jan 15. 99(2):399-408.
  PUBMED    11781219
REFERENCE   2  (bases 1 to 7514)
  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: "Blood. 2002
            Jan 15. 99(2):399-408."
COMMENT     Sequence Label: EF.GFP
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    129..150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(165..170,171..188,189..195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    203..219
                     /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     208..230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     227..243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     227..243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     262..282
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        264..282
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        310..536
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             537..717
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    764..889
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1382..1615
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1800..1844
                     /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        2232..3410
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2463..3401
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3358..3378
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        3427..3445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3509..3518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3515..3517,3518..3520,3521..4234)
                     /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(3515..3517,3518..3520,3521..4231)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3560..3581)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3821..3840)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4168..4189
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4342..4575
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4647..4768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4684..4703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4738..4757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4808..4943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4964..4982)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4992..5009)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4992..5008)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5001..5023)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5150..5605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5237..5256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5447..5468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5631..5735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5736..5804,5805..6596)
                     /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(5954..5973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6767..7355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7256..7275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(7509..7514,1..12)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      121 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
      241 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
      301 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
      361 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
      421 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
      481 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
      541 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      601 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      661 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
      721 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
      781 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
      841 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
      901 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
      961 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1021 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1081 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1141 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1201 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     1261 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1321 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1381 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1441 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1501 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1561 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1621 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1681 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     1741 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     1801 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     1861 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     1921 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     1981 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2041 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
     2101 atcgatcacg agactagcct cgagaagctt gatgacggta tcgataagct ttgcaaagat
     2161 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
     2221 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     2281 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
     2341 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     2401 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     2461 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     2521 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
     2581 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
     2641 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
     2701 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
     2761 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
     2821 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
     2881 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
     2941 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
     3001 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
     3061 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
     3121 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     3181 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     3241 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     3301 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     3361 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattagct
     3421 tggtactaat acgactcact atagggagac ccaagctggc taggtaagct tgatatcgaa
     3481 ttcctgcagc ccgggggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     3541 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3601 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3661 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3721 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3781 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3841 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3901 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3961 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4021 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4081 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4141 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4201 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgtcgacg ggcccgcgga
     4261 attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     4321 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     4381 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     4441 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     4501 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     4561 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     4621 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     4681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4741 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     4801 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4861 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4921 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     4981 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5041 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5101 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5221 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5341 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5461 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5521 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5581 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5641 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5701 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5761 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5821 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5881 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5941 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6001 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6061 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6121 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6181 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6241 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6301 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6361 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6421 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6481 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6541 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6601 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6661 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6721 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6781 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6841 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6901 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6961 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7021 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7081 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7141 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7201 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7261 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7321 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7381 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7441 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7501 ccgagcgcag cgag
//
