LOCUS       pMSCV2.2-mNINJ1-        6936 bp DNA     circular SYN 12-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  AUTHORS   JosAAAA(C) Santos
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..112
                     /label=pBABE-5-fwd
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             166..621
                     /label=mNINJ1
     misc_feature    687..1239
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(719..737)
                     /label=IRES-rev
     CDS             1251..1970
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1293..1309)
                     /label=PEN1-737R
     primer_bind     complement(1359..1377)
                     /label=EGFP-N-rev
     primer_bind     1877..1895
                     /label=EGFP-C-F-31
     primer_bind     1886..1902
                     /label=EGFP-for
     primer_bind     1908..1925
                     /label=EGFP-C-for
     LTR             2023..2537
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2706..2722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2706..2722)
                     /label=M13r
     primer_bind     complement(2720..2739)
                     /label=LacOp-for
     protein_bind    2730..2746
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2754..2784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2754..2773)
                     /label=pUCM13-rev-157
     protein_bind    2799..2820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3108..3696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3867..4727)
                     /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     complement(3917..3936)
                     /label=AmpStop
     primer_bind     4663..4682
                     /label=AmpStart
     promoter        complement(4728..4832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4907..4925
                     /label=QE-for
     primer_bind     complement(4956..4974)
                     /label=GEX3-rev-25
     primer_bind     complement(4984..5003)
                     /label=GEX3-rev
     primer_bind     5254..5273
                     /label=pUCM13-52
     primer_bind     5286..5302
                     /label=M13-40
     primer_bind     5502..5516
                     /label=pET-up
     LTR             5660..6176
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6240..6581
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6648..6936,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ctcgagatcg atgcggcccg ccaccatgga gtcgggcact
      181 gaggagtatg agctcaacgg cgacctgcgc ccgggctccc ccggttcccc cgacgccttg
      241 ccaccccgct ggggtttgcg gaaccggccc atcaatgtaa accattatgc caacaagaag
      301 agcgctgcgg agagcatgct ggccatcgcg ctgctcatgg ccaacgcgtc gcagctgaag
      361 gccgtggtgg agcagggcaa tgatttcgcc ttcttcgtgc cccttgtggt cctcatctct
      421 atctccctcg tgctgcagat aggagtgggc gtgctgctca tcttcctggt caagtatgac
      481 ctcaacaacc cggccaagca cgccaagctg gactttctta acaacctggc cacgggactg
      541 gttttcatca tcgtcgtggt caacatcttc attacggcct tcggggtcca gaagcctgta
      601 atggacgtgg cgccccggca gtaagggccg cgtttaaacg tcgacggtat cgataagctt
      661 gatatcgaat tccgcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
      721 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
      781 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
      841 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
      901 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
      961 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     1021 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     1081 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     1141 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     1201 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca
     1261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1321 acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga
     1381 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1441 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1681 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1741 cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1861 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga taagaattcg
     1981 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     2041 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     2101 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     2161 gatatctgtg gtaagcagtt cctgcccccg gctcagggcc aagaacagat ggtccccgat
     2221 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     2281 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     2341 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     2401 tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     2461 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     2521 cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta
     2581 ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca
     2641 atactcctga aatagttcat tatggacagc gcagaagagc tggggagaat taattcgtaa
     2701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4861 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4921 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4981 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5041 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5101 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5161 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5221 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     5281 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     5341 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5401 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5461 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     5521 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     5581 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     5641 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     5701 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     5761 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     5821 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     5881 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     5941 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6001 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     6061 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     6121 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     6181 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     6241 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     6301 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     6361 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     6421 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     6481 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     6541 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     6601 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     6661 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     6721 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     6781 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     6841 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     6901 gggaagcctt ggcttttgac ccccctccct gggtca
//
