LOCUS       sequence-229586-        7743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control vector, 20nt random sequence between gateway recombination 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7743)
  TITLE     Rehwinkel Lab plasmids
REFERENCE   2  (bases 1 to 7743)
  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: pLenti6.3/EV
FEATURES             Location/Qualifiers
     source          1..7743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2247..2450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2401..2421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2452..2470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2473..2491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2532..2556
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2604..2628)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2681..2722
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2746..3334
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2799..2819)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3217..3228)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3349..3665
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3516..3651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3713..3760
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3779..4177
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4264..4497
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4569..4690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4606..4625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4660..4679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4730..4865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4885..4904)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4886..4904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4914..4931)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4914..4930)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4923..4945)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5072..5527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5159..5178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5369..5390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5553..5657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5658..5726,5727..6518)
                     /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(5876..5895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6689..7277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7178..7197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7431..7448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7565..7586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7601..7606,7607..7624,7625..7631)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7639..7655
                     /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     7644..7666
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7663..7679
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7663..7679
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7698..7718
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7700..7718
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcttacat aacttacggt aaatggcccg cctggctgac
     1981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
     2461 gtgatagaga tctccctatc agtgatagag atcgtcgact agtccagtgt ggtggaattc
     2521 tgcagatatc aacaagtttg tacaaaaaag caggctccga attcgccctt atctacccgc
     2581 gaggtcagac aagggcgaat tcgacccagc tttcttgtac aaagtggttg atatccagca
     2641 cagtggcggc cgctcgagtc tagaggggcc gcggttcgaa ggtaagccta tccctaaccc
     2701 tctcctcggt ctcgattcta cgcgtaccgg ttagtaatga tcgacaatca acctctggat
     2761 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2821 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2881 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2941 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3001 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3061 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3121 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     3181 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3241 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3301 acgagtcgga tctccctttg ggccgcctcc ccgcctggcg atggtaccgg tgtggaaagt
     3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3601 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3661 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     3721 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     3781 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3841 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     3901 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3961 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     4021 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     4081 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     4141 tgaattgctg ccctctggtt atgtgtggga gggctaagca caattcgagc tcggtacctt
     4201 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     4261 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     4321 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     4381 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4441 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     4501 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     4561 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4681 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     4741 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4801 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4861 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4921 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4981 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5041 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5281 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5401 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5521 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5581 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5641 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5701 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5761 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5821 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5881 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5941 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6001 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6061 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6121 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6181 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6241 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6301 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6361 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6421 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6481 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6541 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6601 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6661 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6721 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6781 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6841 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6901 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6961 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7021 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7081 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7141 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7201 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7261 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7321 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7381 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7441 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7501 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7561 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7621 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7681 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     7741 ctt
//
