LOCUS       sequence-244564-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  TITLE     Tsoulfas Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7513)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV-Azurite
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /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    2089..2205
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2228..2531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2532..2735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2686..2706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2732..2756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2811..2820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3123..3142)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3644..4232
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3697..3717)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4115..4126)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4319..4552
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4624..4745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4661..4680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4715..4734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4785..4920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4940..4959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4941..4959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4969..4986)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4969..4985)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4978..5000)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5127..5582
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5214..5233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5424..5445
                     /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"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     2041 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
     2101 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2161 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg
     2221 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2281 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2341 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2401 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     2461 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2521 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     2581 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     2641 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     2701 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     2761 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgcccggtc gccaccatgg
     2821 tgagcaaagg cgaagagctg ttcaccggcg tggtgcccat cctggtggag ctggacggcg
     2881 acgtgaacgg ccacaagttc agcgtgagcg gcgagggcga gggcgacgcc acctacggca
     2941 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctgg
     3001 tgaccaccct gagccacggc gtgcagtgct tcagccggta ccccgaccac atgaagcagc
     3061 acgacttctt caagagcgcc atgcccgagg gctacgtgca ggagcggacc atcttcttca
     3121 aggacgacgg caactacaag acccgggccg aggtgaagtt cgagggcgac accctggtga
     3181 accggatcga gctgaagggc atcgacttca aggaggacgg caacatcctg ggccacaagc
     3241 tggagtacaa cttcaacagc cacaacatct acatcatggc cgacaagcag aagaacggca
     3301 tcaaggtgaa cttcaagatc cggcacaaca tcgaggacgg cagcgtgcag ctggccgacc
     3361 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3421 tgagcaccca gagcgccctg agcaaggacc ccaacgagaa gcgggaccac atggtgctgc
     3481 tggagttccg gaccgccgcc ggcatcaccc acggcatgga cgagctctat aagtaaagcg
     3541 gccaacggat cccaccggtg ctcgagatgc atactagtga attcacgcgt tctagacctg
     3601 cagggtcgac tccggatgat cagggccctg tacagatatc gacaatcaac ctctggatta
     3661 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3721 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3781 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3841 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3901 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3961 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4021 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4081 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4141 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4201 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt acctttaaga
     4261 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4321 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     4381 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4441 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4501 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     4561 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     4621 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4681 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4741 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     4801 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4861 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4921 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     4981 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5041 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5101 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     5161 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5221 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5281 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5341 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5401 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5461 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5521 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5581 ttaggtggca 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 tcttctagtg
     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 aag
//
