LOCUS       sequence-42601-e        8331 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 8331)
  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 8331)
  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: pMA3160
FEATURES             Location/Qualifiers
     source          1..8331
                     /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      3047..3056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3053..4705
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3119..3137)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     4707..4723
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4707..4723
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4772..4792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4777..5106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4957..5092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5120..5719
                     /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(5120..5139)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5616..5636
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6337..6517
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6579..7167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6659..6678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7338..8129,8130..8198))
                     /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     7961..7980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8199..8303)
                     /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 ggagaattct gcagatatcc agcacagtgg cggccgctcg
     3001 agatctgcga tctaagtaag cttggcattc cggtactgtt ggtaaagcca ccatggaaga
     3061 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
     3121 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
     3181 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
     3241 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
     3301 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
     3361 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
     3421 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
     3481 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     3541 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
     3601 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
     3661 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
     3721 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
     3781 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     3841 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
     3901 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
     3961 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     4021 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     4081 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     4141 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     4201 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     4261 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     4321 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     4381 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     4441 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     4501 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     4561 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     4621 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     4681 gaagggcgga aagatcgccg tgtaattcta gaactagtgg atcccccggg ctgcaggaat
     4741 tgatggccgc caccgcggtg gaggaccctg tggaatgtgt gtcagttagg gtgtggaaag
     4801 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4861 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4921 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4981 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5041 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5101 tgcaaaaagc tagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5161 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5221 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5281 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5341 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5401 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5461 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5521 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5581 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5641 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5701 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     5761 acgaccccat gcatcgatca attcgatggg ggatcaattc gattaagtcg agggtcgatc
     5821 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     5881 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     5941 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6001 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6061 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     6121 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     6181 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     6241 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     6301 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     6361 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6421 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6481 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     6541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7261 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7321 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7381 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7441 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7501 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7561 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7621 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7681 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7741 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7801 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7861 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7921 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7981 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8041 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8101 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8161 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
