LOCUS       sequence-422314-        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for MLV Gag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  AUTHORS   Najia MA, Borrajo J, Le A, Tsai F, Huang JY, Griffith LG, Daley GQ, 
            Blainey PC
  TITLE     Live-cell transcriptomics with engineered virus-like particles
  JOURNAL   bioRxiv 2024.10.01.616098
REFERENCE   2  (bases 1 to 7053)
  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: "bioRxiv 
            2024.10.01.616098"
COMMENT     Sequence Label: pGag
FEATURES             Location/Qualifiers
     source          1..7053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     662..686
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          799..1274
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             1332..1748
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1678..1700
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1716..1732
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2928..2984
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2985..3683
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFAR
                     TFSLRDGGYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAF
                     KTPDADAGEE"
     primer_bind     complement(3702..3719)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3708..3915
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4059..4453
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(4785..4803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4871..4975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4976..5044,5045..5836)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5194..5213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     6372..6391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat
       61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      721 atagaagaca ccgggaccga tccagcctcc cctcgaagct tacatgtggt accgagctcg
      781 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
      841 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
      901 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
      961 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
     1021 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
     1081 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
     1141 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
     1201 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
     1261 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
     1321 aagaattcat gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1381 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1441 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1501 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1561 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1621 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1681 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1741 taggcgccaa acctaaacct caagttcttt ctgacagtgg ggggccgctc atcgacctac
     1801 ttacagaaga ccccccgcct tatagggacc caagaccacc cccttccgac agggacggaa
     1861 atggtggaga agcgacccct gcgggagagg caccggaccc ctccccaatg gcatctcgcc
     1921 tacgtgggag acgggagccc cctgtggccg actccactac ctcgcaggca ttccccctcc
     1981 gcgcaggagg aaacggacag cttcaatact ggccgttctc ctcttctgac ctttacaact
     2041 ggaaaaataa taacccttct ttttctgaag atccaggtaa actgacagct ctgatcgagt
     2101 ctgtcctcat cacccatcag cccacctggg acgactgtca gcagctgttg gggactctgc
     2161 tgaccggaga agaaaaacaa cgggtgctct tagaggctag aaaggcggtg cggggcgatg
     2221 atgggcgccc cactcaactg cccaatgaag tcgatgccgc ttttcccctc gagcgcccag
     2281 actgggatta caccacccag gcaggtagga accacctagt ccactatcgc cagttgctcc
     2341 tagcgggtct ccaaaacgcg ggcagaagcc ccaccaattt ggccaaggta aaaggaataa
     2401 cacaagggcc caatgagtct ccctcggcct tcctagagag acttaaggaa gcctatcgca
     2461 ggtacactcc ttatgaccct gaggacccag ggcaagaaac taatgtgtct atgtctttca
     2521 tttggcagtc tgccccagac attgggagaa agttagagag gttagaagat ttaaaaaaca
     2581 agacgcttgg agatttggtt agagaggcag aaaagatctt taataaacga gaaaccccgg
     2641 aagaaagaga ggaacgtatc aggagagaaa cagaggaaaa agaagaacgc cgtaggacag
     2701 aggatgagca gaaagagaaa gaaagagatc gtaggagaca tagagagatg agcaagctat
     2761 tggccactgt cgttagtgga cagaaacagg atagacaggg aggagaacga aggaggtccc
     2821 aactcgatcg cgaccagtgt gcctactgca aagaaaaggg gcactgggct aaagattgtc
     2881 ccaagaaacc acgaggacct cggggaccaa gaccgcaggg atccggcgca acaaacttct
     2941 ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgatggag agcgacgaga
     3001 gcggcctgcc cgccatgaag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt
     3061 tcgagctggt gggcggcgga gagggcaccc ccgagcaggg ccgcatgacc aacaagatga
     3121 agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg
     3181 gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca
     3241 acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg
     3301 tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca
     3361 ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg
     3421 tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca
     3481 gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg
     3541 ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg
     3601 agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccgg
     3661 atgcagatgc cggtgaagaa tagctagagc tcgctgatca gcctcgactg tgccttctag
     3721 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3781 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3841 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     3901 caggcatgct ggggacatgg cgcgttcaac gctctcaaaa ccccttaaaa ataaggttaa
     3961 cccgcgaggc cccctaagaa ttcaccccac cagtgcaggc tgcctatcag aaagtggtgg
     4021 ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt gctgtccaat
     4081 ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga tattatgaag
     4141 ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc aatgatgtat
     4201 ttaaattatt tctgaatatt ttactaaaaa gggaatgtgg gaggtcagtg catttaaaac
     4261 ataaagaaat gaagagctag ttcaaacctt gggaaaatac actatatctt aaactccatg
     4321 aaagaaggtg aggctgcaaa cagctaatgc acattggcaa cagcccctga tgcctatgcc
     4381 ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta aagttttgct
     4441 atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt ttcactaccc
     4501 atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct tagagatacc
     4561 acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc ctcgcctggc
     4621 cactcagcct tagttgtctc tgttgtctta tagaggtcta cttgaagaag gaaaaacagg
     4681 gggcatggtt tgactgtcct gtgagccctt cttccctgcc tcccccactc acagtgaccc
     4741 ggaatccctc gacatggcag tctagaattc ttgaagacga aagggcctcg tgatacgcct
     4801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     5521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5881 acttcatttt taatttaaaa ggatctaggc tgctgcttgc aaacaaaaaa accaccgcta
     5941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6001 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     6061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6481 aacggagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     6541 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     6601 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     6661 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     6721 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     6781 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     6841 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     6901 agaatcataa tggggaaggc catccagcct cgcgtcgggg agctttttgc aaaagcctag
     6961 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     7021 ctctgcataa ataaaaaaaa ttagtcagcc atg
//
