LOCUS       sequence-266188-        7702 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 7702)
  AUTHORS   Gaiano N, Nye JS, Fishell G
  TITLE     Radial glial identity is promoted by Notch1 signaling in the murine 
            forebrain.
  JOURNAL   Neuron. 2000 May . 26(2):395-404.
  PUBMED    10839358
REFERENCE   2  (bases 1 to 7702)
  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: "Neuron. 
            2000 May . 26(2):395-404."
COMMENT     Sequence Label: pCLE
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    823..1022
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1223..1639
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1569..1591
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1607..1623
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2181..2733
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2325..2342)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2552..2571
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2745..4244
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="LLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPA
                     QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
                     VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
                     TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
                     MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHL
                     MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRA
                     YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKAR
                     DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
                     GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     LTR             4810..5279
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5409..5426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5580..6168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5660..5679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6328..7119,7120..7188))
                     /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     6951..6970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7189..7293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7361..7379
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7417..7439)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7539..7558
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
      601 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
      661 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      721 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
      781 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
      841 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
      901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1621 cactccttct ctaggcgccg gccggatcga gcagggggat catctaatca agcacaaata
     1681 aggggcgtgt aacacaaaag ccagcgaccc tttccaatgc aaatcaaact tgcaattctt
     1741 tgccgttttt atcatttaag tgtcggctta aggtccacta tcagatgtaa acagccttat
     1801 ctaacaaagg tatcattaca ttctgaaatt ctcaggcatg caagctagct tatgacgcac
     1861 tagggagtgc cacccttcct ttcgccctaa cttcgtgata actcgcgcgt ttcactcaac
     1921 agctgcatcc gccctagtgc tactgggagt tgtagtatac aagacgctta caggctgaat
     1981 gttctgtcaa gaccccgcct ctagcacttt gggaattctg gacttgatga tgtcatggtt
     2041 aatccccgcc cagtagaggc ggctatataa agggtggtta aggcccggtt cgctctcttc
     2101 ctcaccgggt ctgcggcgag ttctagctga agctgatccc agtgtggtgg taccagatct
     2161 ggtaggaatt ccgcccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     2221 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2281 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2341 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2401 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2461 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2521 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2581 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     2641 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2701 cgtggttttc ctttgaaaaa cacgatgata atatggggtc gaggctgctg ctgctgctgc
     2761 tgctgggcct gaggctacag ctctccctgg gcatcatccc agttgaggag gagaacccgg
     2821 acttctggaa ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac
     2881 agacagccgc caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga
     2941 cagctgccag gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg
     3001 ccatggaccg cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc
     3061 cagacagtgg agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca
     3121 ttggcttgag tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca
     3181 tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac
     3241 gagtgcagca cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact
     3301 cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc
     3361 tcatctccaa catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca
     3421 tgggaacccc agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg
     3481 ggaagaatct ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc
     3541 gcactgagct catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg
     3601 agcctggaga catgaaatac gagatccacc gagactccac actggacccc tccctgatgg
     3661 agatgacaga ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg
     3721 tggagggtgg tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg
     3781 agacgatcat gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca
     3841 cgctgagcct cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc
     3901 gagggagctc catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg
     3961 tcctcctata cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta
     4021 ccgagagcga gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag
     4081 agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc
     4141 acggcgtgca ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc
     4201 cctacaccgc ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccggggc
     4261 ggtccgtggt ccccgcgttg cttcctctgc tggccgggac cctgctgctg ctggagacgg
     4321 ccactgctcc ctgagtgtcc cgtccctggg gctcctgctt ccccatcccg gagttctcct
     4381 gctccccacc tcctgtcgtc ctgcctggcc tccagcccga gtcgtcatcc ccggagtccc
     4441 tatacagagg tcctgccatg gaaccttccc ctccccgtgc gctctgggga ctgagcccat
     4501 gacaccaaac ctgccccttg gctgctctcg gactccctac cccaacccca gggactgcag
     4561 gttgtgccct gtggctgcct gcaccccagg aaaggagggg gctcaggcca tccagccacc
     4621 acctacagcc cagtggggtc gacccgataa aataaaagat tttatttagt ctccagaaaa
     4681 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4741 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     4801 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4861 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     4921 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     4981 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     5041 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     5101 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     5161 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     5221 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     5281 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     5341 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6241 atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag
     6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7321 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7381 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     7441 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     7501 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     7561 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7621 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7681 ttcgctatta cgccagcgcg tt
//
