LOCUS       sequence-251302-        6422 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 6422)
  TITLE     pBabe-IRES-EGFP plasmids
REFERENCE   2  (bases 1 to 6422)
  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"
COMMENT     Sequence Label: pBabe puro IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..533
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     463..485
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     501..517
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(611..1185,1186..1188,1189..1197)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(778..795)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1005..1024
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1198..1200,1201..1203,1204..1917)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1198..1200,1201..1203,1204..1914)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1243..1264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1504..1523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1851..1872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1925..1945)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1930..2259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2110..2245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2269..2868
                     /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(2269..2288)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2765..2785
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3091..3560
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3690..3707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3861..4449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3941..3960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4609..5400,5401..5469))
                     /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     5232..5251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5470..5574)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5605..6074
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6139..6338
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
       61 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
      121 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      181 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      241 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
      301 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
      361 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
      421 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
      481 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggccgga
      541 tccgggctcg agcccgaatt cgggagatct cccatcgatg gggtcgacgg taccgcgggc
      601 ccgggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
      661 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      721 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
      781 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
      841 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
      901 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
      961 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1021 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     1081 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     1141 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     1201 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1261 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1321 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1381 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1441 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1501 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1561 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1621 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1681 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1741 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1801 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1861 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagcc
     1921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2161 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac
     2281 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     2341 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     2401 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     2461 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     2521 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     2581 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     2641 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     2701 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     2761 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     2821 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     2881 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgcgata aaataaaaga
     2941 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     3001 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     3061 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     3121 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     3181 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     3241 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     3301 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     3361 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     3421 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     3481 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     3541 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     3601 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     3661 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3721 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3781 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4261 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4501 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     4561 cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5521 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5581 tccccgaaaa gtgccacctg cagcctgaat atgggccaaa caggatatct gtggtaagca
     5641 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     5701 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     5761 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     5821 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     5881 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     5941 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     6001 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     6061 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
     6121 gacccaccac cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt
     6181 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
     6241 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
     6301 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
     6361 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     6421 gt
//
