LOCUS       sequence-39434-e        7191 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 7191)
  AUTHORS   Alexeyev MF, Fayzulin R, Shokolenko IN, Pastukh V
  TITLE     A retro-lentiviral system for doxycycline-inducible gene expression 
            and gene knockdown in cells with limited proliferative capacity.
  JOURNAL   Mol Biol Rep. 2010 Apr . 37(4):1987-91.
  PUBMED    19655272
REFERENCE   2  (bases 1 to 7191)
  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: "Mol Biol 
            Rep. 2010 Apr . 37(4):1987-91."
COMMENT     Sequence Label: pMA2706
FEATURES             Location/Qualifiers
     source          1..7191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1035..1160
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1657..1890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2075..2119
                     /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    2413..2530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(2629..2645)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2630..2646)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        2645..2959
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2653..2671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2689..2707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2724..2742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2760..2778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2796..2814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2831..2849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2867..2885
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2932..2956
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2995..3004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3001..3558
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     primer_bind     3567..3583
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3567..3583
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3632..3652)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3637..3966
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3817..3952
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3980..4579
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3980..3999)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4476..4496
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5197..5377
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5519..5538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6198..6989,6990..7058))
                     /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     6821..6840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7059..7163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
      241 gcgttgacat tgattattga ctagctagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tagaccagat ctgagcctgg
      841 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      901 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      961 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     1021 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1081 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1141 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1201 ttcggttaag gccaggggga aagaaaaaat ataaataaac atatagtatg ggcaagcagg
     1261 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     1321 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1381 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1441 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1501 ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1561 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1621 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1681 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1741 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1801 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1861 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1921 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1981 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     2041 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     2101 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     2161 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2221 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2281 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2341 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattcgac aaatggcagt
     2401 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2461 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2521 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag taccgggccg
     2581 gcccgctcta gctagacgta ttaccgccag atcccccatc aagcttatcg ataccgtcga
     2641 cctcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
     2701 atagagaacg atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
     2761 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat
     2821 gtcgagttta tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     2881 agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
     2941 agtgaaccgt cagatcgcct ggagaattcg agctcggtac cgagctcgga tccagccacc
     3001 atgggagtca aagttctgtt tgccctgatc tgcatcgctg tggccgaggc caagcccacc
     3061 gagaacaacg aagacttcaa catcgtggcc gtggccagca acttcgcgac cacggatctc
     3121 gatgctgacc gcgggaagtt gcccggcaag aagctgccgc tggaggtgct caaagagatg
     3181 gaagccaatg cccggaaagc tggctgcacc aggggctgtc tgatctgcct gtcccacatc
     3241 aagtgcacgc ccaagatgaa gaagttcatc ccaggacgct gccacaccta cgaaggcgac
     3301 aaagagtccg cacagggcgg cataggcgag gcgatcgtcg acattcctga gattcctggg
     3361 ttcaaggact tggagcccat ggagcagttc atcgcacagg tcgatctgtg tgtggactgc
     3421 acaactggct gcctcaaagg gcttgccaac gtgcagtgtt ctgacctgct caagaagtgg
     3481 ctgccgcaac gctgtgcgac ctttgccagc aagatccagg gccaggtgga caagatcaag
     3541 ggggccggtg gtgactaagc ggccgctcta gaactagtgg atcccccggg ctgcaggaat
     3601 tgatggccgc caccgcggtg gaggaccctg tggaatgtgt gtcagttagg gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3781 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3841 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3901 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3961 tgcaaaaagc tagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4021 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4081 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4141 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4201 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4261 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4321 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4381 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4441 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4501 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4561 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     4621 acgaccccat gcatcgatca attcgatggg ggatcaattc gattaagtcg agggtcgatc
     4681 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     4741 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     4801 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     4861 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     4921 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     4981 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     5041 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     5101 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     5161 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     5221 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5281 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5341 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
