LOCUS       sequence-7285-ex        7819 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7819)
  AUTHORS   Naviaux RK, Costanzi E, Haas M, Verma IM
  TITLE     The pCL vector system: rapid production of helper-free, high-titer, 
            recombinant retroviruses.
  JOURNAL   J Virol. 1996 Aug . 70(8):5701-5.
  PUBMED    8764092
REFERENCE   2  (bases 1 to 7819)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Virol. 
            1996 Aug . 70(8):5701-5."
COMMENT     Sequence Label: pCLXSN
FEATURES             Location/Qualifiers
     source          1..7819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             589..764
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    827..1026
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1227..1643
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1573..1595
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1611..1627
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1685..1705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1690..2019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1870..2005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2079..2873
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2133..2152)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2743..2762
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             2933..3526
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          3873..3938
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4067..4087
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4359..4480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4396..4415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4450..4469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5525..5542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5696..6284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5776..5795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6753..7129)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     7076..7095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7314..7418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7486..7504
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     7664..7683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatgtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc
      601 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
      661 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      721 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      781 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      841 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      901 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      961 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     1021 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     1081 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     1141 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     1201 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1261 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1321 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1381 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1441 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1501 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1561 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1621 ccctcactcc ttctctaggc gccggaattg cggccgcaat tcgttaactc gaggatccgg
     1681 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1741 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1861 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1921 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     1981 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgggct gcaggtcgag
     2041 gcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2101 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2161 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2221 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2281 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2341 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2401 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2461 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2521 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2581 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2641 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2701 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2761 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2821 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     2881 tctggggttc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     2941 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     3001 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     3061 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3121 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3181 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     3241 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3301 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     3361 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     3421 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     3481 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg
     3541 ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc
     3601 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3661 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3721 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatacct
     3781 agctaggtag ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     3841 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     3901 gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg
     3961 aatgggagca gtggtccaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc
     4021 ctctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag
     4081 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt
     4141 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact
     4201 gctatacaag aaaattatgg aaaaatattt gatgtatagt gccttgacta gagatcataa
     4261 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4321 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4441 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctga tcccgccatg
     4501 gtatcaacgc catatttcta tttacagtag ggacctcttc gttgtgtagg taccgctgta
     4561 ttcctaggga aatagtagag gcaccttgaa ctgtctgcat cagccatata gcccccgctg
     4621 ttcgacttac aaacacaggc acagtactga caaacccata cacctcctct gaaataccca
     4681 tagttgctag ggctgtctcc gaactcatta caccctccaa agtcagagct gtaatttcgc
     4741 catcaagggc agcgagggct tctccagata aaatagcttc tgccgagagt cccgtaaggg
     4801 tagacacttc agctaatccc tcgatgaggt ctactagaat agtcagtgcg gctcccattt
     4861 tgaaaattca cttacttgat cagcttcaga agatggcgga gggcctccaa cacagtaatt
     4921 ttcctcccga ctcttaaaat agaaaatgtc aagtcagtta agcaggaagt ggactaactg
     4981 acgcagctgg ccgtgcgaca tcctctttta attagttgct aggcaacgcc ctccagaggg
     5041 cgtgtggttt tgcaagagga agcaaaagcc tctccaccca ggcctagaat gtttccaccc
     5101 aatcattact atgacaacag ctgttttttt tagtattaag cagaggccgg ggacccctgg
     5161 cccgcttact ctggagaaaa aaaacattgt agaggcttcc agaggcaact tgtcaaaaca
     5221 ggactgcttc tatttctgtc acactgtctg gccctgtcac aaggtccagc acctccatac
     5281 cccctttaat aagcagtttg ggaacgggtg cgggtcttac tccgcccatc cgcccctaac
     5341 tccgcccagt tccgcccatt ctccgcccca tgctgactaa ttttttttat ttatgcagag
     5401 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5461 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5521 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5581 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5641 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6121 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     6781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaactc aaccaagtca
     7021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7441 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7501 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     7561 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     7621 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     7681 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     7741 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     7801 gcctcttcgc tattacgcc
//
