LOCUS       sequence-323904-        7547 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 7547)
  TITLE     TRONO lab plasmids
REFERENCE   2  (bases 1 to 7547)
  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"
COMMENT     Sequence Label: pAlbumin
FEATURES             Location/Qualifiers
     source          1..7547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(182..250,251..1042)
                     /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(400..419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1213..1801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1702..1721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2114..2131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2248..2269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2284..2289,2290..2307,2308..2314)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2322..2338
                     /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     2327..2349
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2346..2362
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2346..2362
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2381..2401
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2383..2401
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2429..2655
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2656..2836
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2883..3008
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3501..3734
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3919..3963
                     /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    4230..4347
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4393..4903
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4822..4840
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      4919..4928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4925..4927,4928..4930,4931..5644)
                     /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(4925..4927,4928..4930,4931..5641)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4970..4991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5231..5250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5578..5599
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5660..6248
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5713..5733)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6131..6142)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6335..6568
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6640..6761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6677..6696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6731..6750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6801..6936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6956..6975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6957..6975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6985..7002)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6985..7001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6994..7016)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(7143..7547,1..51)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7230..7249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7440..7461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
       61 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      181 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      241 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      301 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      361 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      421 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      481 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
      541 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
      601 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
      661 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
      721 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
      781 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
      841 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
      901 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
      961 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1021 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1081 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1141 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1201 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1261 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1321 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1381 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1441 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1501 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1561 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1621 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1681 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1741 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1801 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacaa
     1861 gcttggggtt gctgtcatct cttgtgggct gtaatcatcg tctaggctta agagtaatat
     1921 tgcaaaacct gtcatgccca cacaaatctc tccctggcat tgttgtcttt gcagatgtca
     1981 gtgaaagaga accagcagct cccatgagtc ccaagctatg ttctttcctg cgttatcccc
     2041 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2101 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     2161 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2221 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     2281 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     2341 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     2401 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     2461 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     2521 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     2581 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     2641 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     2701 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     2761 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     2821 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     2881 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     2941 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     3001 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     3061 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     3121 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     3181 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     3241 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     3301 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     3361 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     3421 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     3481 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     3541 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     3601 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     3661 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     3721 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     3781 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     3841 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     3901 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     3961 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     4021 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     4081 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     4141 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     4201 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
     4261 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     4321 aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga gaagcttgat
     4381 atcgaattcc acggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg
     4441 acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac
     4501 attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc
     4561 ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg
     4621 gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg
     4681 agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg
     4741 cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg
     4801 ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc
     4861 agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggtcgc
     4921 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4981 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     5041 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     5101 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     5161 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     5221 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     5281 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     5341 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     5401 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     5461 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     5521 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     5581 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5641 gtaaagcggc cgcgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5701 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5761 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5821 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5881 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5941 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6001 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6061 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6121 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6181 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6241 ctccccgcct ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta
     6301 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6361 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     6421 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6481 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6541 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6601 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     6781 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6841 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6901 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     6961 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     7021 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     7081 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     7141 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     7201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     7261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     7321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     7381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     7441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     7501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctga
//
