LOCUS       sequence-204864-        7086 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 7086)
  AUTHORS   Dyer MA, Cepko CL
  TITLE     p27Kip1 and p57Kip2 regulate proliferation in distinct retinal 
            progenitor cell populations.
  JOURNAL   J Neurosci. 2001 Jun 15. 21(12):4259-71.
  PUBMED    11404411
REFERENCE   2  (bases 1 to 7086)
  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: "J Neurosci.
            2001 Jun 15. 21(12):4259-71."
COMMENT     Sequence Label: pGFP-E p27 (CC#631)
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    257..614
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             679..1095
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1025..1047
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1063..1079
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1105..1479
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1417..1433
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1554..1577
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2184..2201
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(2293..2867,2868..2870,2871..2879)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2460..2477)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2687..2706
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2880..2882,2883..2885,2886..3599)
                     /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(2880..2882,2883..2885,2886..3596)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2925..2946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3186..3205)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3533..3554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3785..4254
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4384..4401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4555..5143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4635..4654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5267..5335,5336..6127)
                     /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(5485..5504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     LTR             join(6809..7086,1..192)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
       61 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
      121 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
      181 gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga
      241 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
      301 ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc
      361 ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca
      421 gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga
      481 ctctttggtg cacccccctt agaggaggga tatgtggttc tggtaggaga cgagaaccta
      541 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg
      601 cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
      661 tctgaaaata tgggcccggg ccagactgtt accactccct taagtttgac cttaggtcac
      721 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
      781 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
      841 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
      901 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
      961 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     1021 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     1081 ccttctctag gcgcccccat atggccatat gagatcttat atggggcacc cccgcccctt
     1141 gtaaacttcc ctgaccctga catgacaaga gttactaaca gcccctctct ccaagctcac
     1201 ttacaggctc tctacttagt ccagcacgaa gtctggagac ctctggcggc agcctaccaa
     1261 gaacaactgg accgaccggt ggtacctcac ccttaccgag tcggcgacac agtgtgggtc
     1321 cgccgacacc agactaagaa cctagaacct cgctggaaag gaccttacac agtcctgctg
     1381 accaccccca ccgccctcaa agtagacggc atcgcagctt ggatacacgc cgcccacgtg
     1441 aaggctgccg accccggggg tggaccatcc tctagactgc cggatccatt taaattcgaa
     1501 ttccggtcga gcagctgaag cttgcatgcc tgcaggtcga ctctagaacc atggactaca
     1561 aggacgacga tgacaaggcg gccgcatcaa acgtgagagt gtctaacggg agcccgagcc
     1621 tggagcggat ggacgccaga caagcggatc accccaagcc ttccgcctgc agaaatctct
     1681 tcggcccggt caatcatgaa gaactaaccc gggacttgga gaagcactgc cgggatatgg
     1741 aagaagcgag tcagcgcaag tggaatttcg actttcagaa tcataagccc ctggagggca
     1801 gatacgaatg gcaggaggtg gagaggggca gcttgcccga gttctactac aggcccccgc
     1861 gcccccccaa gagcgcctgc aaggtgctgg cgcaggagag ccaggatgtc agcgggagcc
     1921 gccaggcggt gcctttaatt gggtctcagg caaactctga ggaccggcat ttggtggacc
     1981 aaatgcctga ctcgtcagac aatcaggctg ggttagcgga gcagtgtcca gggatgagga
     2041 agcgacctgc tgcagaagat tcttcttcgc aaaacaaaag ggccaacaga acagaagaaa
     2101 atgtttcaga cggttccccg aacgctggca ctgtggagca gacgcccaag aagcccggcc
     2161 ttcgacgcca gacggcggcc gcacatcatc atcatcatca ttaatctaga ggatccccgg
     2221 gtaccgagct cgaattcctg caggcctcga gggccggcgc gccgcggccg ctacgtaaat
     2281 tccgcccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     2341 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2401 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2461 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2521 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2581 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2641 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2701 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2761 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2821 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     2881 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2941 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3001 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3061 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3121 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3181 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3241 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3301 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3361 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3421 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3481 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3541 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     3601 gcgccagcac agtggtcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     3661 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     3721 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     3781 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     3841 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     3901 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     3961 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     4021 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     4081 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     4141 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     4201 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     4261 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4381 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5221 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5281 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5341 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5401 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5461 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5521 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5581 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5641 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5701 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5761 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5821 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5881 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5941 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6001 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6061 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6121 ttggtaactg tcagaccaag tttactcata tatactttag attgatttgc ggccggccgc
     6181 aaaaggatct aggtgaagat cctttttgat aatctcatga gcggatacat atttgaatgt
     6241 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccaccagct
     6301 ttgctcttag gagtttccta atacatccca aactcaaata tataaagcat ttgacttgtt
     6361 ctatgcccta gggggcgggg ggaagctaag ccagcttttt ttaacattta aaatgttaat
     6421 tccattttaa atgcacagat gtttttattt cataagggtt tcaatgtgca tgaatgctgc
     6481 aatattcctg ttaccaaagc tagtataaat aaaaatagat aaacgtggaa attacttaga
     6541 gtttctgtca ttaacgtttc cttcctcagt tgacaacata aatgcgctgc tgagaagcca
     6601 gtttgcatct gtcaggatca atttcccatt atgccagtca tattaattac tagtcaatta
     6661 gttgattttt atttttgaca tatacatgtg aaagacccca cctgtaggtt tggcaagcta
     6721 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agaaaagttc
     6781 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     6841 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     6901 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     6961 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     7021 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     7081 tcgcgc
//
