LOCUS       sequence-304846-        7629 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 7629)
  TITLE     Cookson Lab Plasmids
REFERENCE   2  (bases 1 to 7629)
  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: pLenti6-DJ1-V5-WT
FEATURES             Location/Qualifiers
     source          1..7629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    505..526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(541..546,547..564,565..571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..595
                     /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     584..606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     603..619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     603..619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     638..658
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        640..658
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        686..912
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             913..1093
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1140..1265
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1758..1991
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2176..2220
                     /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"
     enhancer        2499..2802
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2803..3006
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2957..2977
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3003..3027
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3123..3147
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3745..3769)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3822..3863
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3905..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4282..4329
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4348..4746
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4833..5066
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5138..5259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5175..5194)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5229..5248
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5299..5434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5454..5473)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5455..5473)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5483..5500)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5483..5499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5492..5514)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5641..6096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5728..5747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5938..5959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6122..6226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6227..6295,6296..7087)
                     /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(6445..6464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7258..7629,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      601 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      661 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
      721 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
      781 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
      841 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
      901 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      961 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     1021 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     1081 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     1141 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     1201 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     1261 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     1321 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     1381 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     1441 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     1501 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1561 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     1621 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1681 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1741 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1801 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1861 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1921 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1981 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     2041 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     2101 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     2161 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     2221 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     2281 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     2341 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2401 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2461 aacggatctc gacggtatcg ataagcttgg gagttccgcg ttacataact tacggtaaat
     2521 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2581 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2641 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2701 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2761 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2821 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2881 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2941 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     3001 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     3061 catagaagac accgactcta gaggatccac tagtccagtg tggtggaatt ctgcagatat
     3121 caacaagttt gtacaaaaaa gcaggctccg aattcgccct tatggcttcc aaaagagctc
     3181 tggtcatcct ggctaaagga gcagaggaaa tggagacggt catccctgta gatgtcatga
     3241 ggcgagctgg gattaaggtc accgttgcag gcctggctgg aaaagaccca gtacagtgta
     3301 gccgtgatgt ggtcatttgt cctgatgcca gccttgaaga tgcaaaaaaa gagggaccat
     3361 atgatgtggt ggttctacca ggaggtaatc tgggcgcaca gaatttatct gagtctgctg
     3421 ctgtgaagga gatactgaag gagcaggaaa accggaaggg cctgatagcc gccatctgtg
     3481 caggtcctac tgctctgttg gctcatgaaa taggttttgg aagtaaagtt acaacacacc
     3541 ctcttgctaa agacaaaatg atgaatggag gtcattacac ctactctgag aatcgtgtgg
     3601 aaaaagacgg cctgattctt acaagccggg ggcctgggac cagcttcgag tttgcgcttg
     3661 caattgttga agccctgaat ggcaaggagg tggcggctca agtgaaggct ccacttgttc
     3721 ttaaagacaa gaagggcgaa ttcgacccag ctttcttgta caaagtggtt gatatccagc
     3781 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     3841 ctctcctcgg tctcgattct acgcgtaccg gttagtaatg agtttggaat taattctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4261 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     4321 taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc
     4381 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     4441 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     4501 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     4561 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     4621 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     4681 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     4741 ggctaagcac aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     4801 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     4861 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     4921 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4981 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     5041 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     5101 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     5161 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5221 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     5281 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5341 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5401 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     5461 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5521 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5581 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     5641 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5701 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5761 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5821 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     5881 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5941 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6001 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6061 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     6121 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6181 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6241 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6301 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6361 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6421 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6481 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6541 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6601 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6661 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6721 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6781 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6841 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6901 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6961 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7021 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7081 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7141 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7201 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7261 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7321 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7381 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7441 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7501 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7561 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7621 caccgaact
//
