LOCUS       sequence-328004-        5861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of CENP-R uORF-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5861)
  TITLE     CENP-R uORF-1 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5861)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAN68
FEATURES             Location/Qualifiers
     source          1..5861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1491..1508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1518..1541
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1542..1565
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1575..1577,1578..1580,1581..2294)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTATGITLGMDELYK"
     primer_bind     complement(1620..1641)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1881..1900)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2228..2249
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2439..2459)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2444..2773
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2624..2759
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2781..3179
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3348..3817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3947..3964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4118..4706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4198..4217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4866..5657,5658..5726))
                     /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     5489..5508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5727..5831)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ctcgagatgc cgtggcgttt
     1381 cttgtggcgt atcttctgtt tccgcaagct gctgggttcg ttctattcag cggctgtggt
     1441 actttcgcgc atttctgaat gtctgttgaa ggaccacccg cggcagcagc catcatcatc
     1501 atcatcacag cagcggcctg gaagttctgt tccaggggcc cctggaagtt ctgttccagg
     1561 ggccctccgg agggatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1621 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1681 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1741 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1801 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1861 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1921 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1981 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2041 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2101 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2161 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2221 gcgatcacat ggtcctgctg gagttcgtga ccgccaccgg gatcactctt ggcatggacg
     2281 agctgtacaa gtaaagatcc accggatcta gataactgat cataatcagc cataccacat
     2341 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2401 aaatgaatgc aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg
     2461 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2521 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2581 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2641 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2701 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2761 aggcttttgc aaaaagcttc atggccaagc ctttgtctca agaagaatcc accctcattg
     2821 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     2881 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     2941 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     3001 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     3061 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     3121 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     3181 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3241 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     3301 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     3361 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3421 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3481 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3541 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3601 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3661 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3721 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3781 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     3841 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     3901 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3961 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4021 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4741 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4801 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag c
//
