LOCUS       sequence-39435-e        6686 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 6686)
  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 6686)
  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: pMA2780
FEATURES             Location/Qualifiers
     source          1..6686
                     /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"
     promoter        2649..2963
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2657..2675
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2693..2711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2728..2746
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2764..2782
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2800..2818
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2835..2853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2871..2889
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2936..2960
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2988..3004)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3062..3078
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3062..3078
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3127..3147)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3132..3461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3312..3447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3475..4074
                     /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(3475..3494)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3971..3991
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4692..4872
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4934..5522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5014..5033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5693..6484,6485..6553))
                     /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     6316..6335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6554..6658)
                     /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 cctcgatcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
     2701 agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt
     2761 tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
     2821 gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     2881 tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc
     2941 gtttagtgaa ccgtcagatc gcctggagaa ttcgatatca agcttatcga taccgtcgac
     3001 ctcgaggggg ggcccggtac cgaggacgcg tcaattcgag ctccaccgcg gtggcggccg
     3061 ctctagaact agtggatccc ccgggctgca ggaattgatg gccgccaccg cggtggagga
     3121 ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3301 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3361 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3421 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctagct taccatgacc
     3481 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3541 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3601 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3661 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3721 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3781 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3841 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3901 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3961 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4021 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     4081 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gatcaattcg
     4141 atgggggatc aattcgatta agtcgagggt cgatcgacct cgatcgagac ctagaaaaac
     4201 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
     4261 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
     4321 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     4381 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
     4441 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
     4501 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
     4561 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
     4621 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
     4681 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     4741 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     4801 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     4861 aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6661 catttccccg aaaagtgcca cctgac
//
