LOCUS       sequence-240252-        9716 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 9716)
  AUTHORS   Dang L, Yoon K, Wang M, Gaiano N
  TITLE     Notch3 signaling promotes radial glial/progenitor character in the 
            mammalian telencephalon.
  JOURNAL   Dev Neurosci. 2006;28(1-2):58-69.
  PUBMED    16508304
REFERENCE   2  (bases 1 to 9716)
  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: "Dev 
            Neurosci."; date: "2006"; volume: "28(1-2)"; pages: "58-69"
COMMENT     Sequence Label: pCLE NICD3
FEATURES             Location/Qualifiers
     source          1..9716
                     /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"
     regulatory      2167..2176
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4195..4747
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4339..4356)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4566..4585
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4759..6258
                     /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             6824..7293
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7423..7440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7594..8182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7674..7693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8342..9133,9134..9202))
                     /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     8965..8984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9203..9307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9375..9393
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9431..9453)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9553..9572
                     /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 cgaccaccat ggtggcccgg cgcaagcgcg agcacagcac cctctggttc cctgagggct
     2221 tctcactgca caaggacgtg gcctctggtc acaagggccg gcgggaaccc gtgggccagg
     2281 acgcgctggg catgaagaac atggccaagg gtgagagcct gatgggggag gtggccacag
     2341 actggatgga cacagagtgc ccagaggcca agcggctaaa ggtagaggag ccaggcatgg
     2401 gggctgagga ggctgtggat tgccgtcagt ggactcaaca ccatctggtt actgctgaca
     2461 tccgcgtggc accagccatg gcactgacac caccacaggg cgacgcagat gctgatggca
     2521 tggatgtcaa tgtgcgtggc ccagatggct tcaccccgct aatgctggct tccttctgtg
     2581 ggggggctct ggagccaatg ccaactgaag aggatgaggc agatgacaca tcagctagca
     2641 tcatctccga cctgatctgc cagggggctc agcttggggc acggactgac cgtactggcg
     2701 agactgcttt gcacctggct gcccgttatg cccgtgctga tgcagccaag cggctgctgg
     2761 atgctggggc agacaccaat gcccaggacc actcaggccg cactcccctg cacacagctg
     2821 tcacagccga tgcccagggt gtcttccaga ttctcatccg aaaccgctct acagacttgg
     2881 atgcccgcat ggcagatggc tcaacggcac tgatcctggc ggcccgcctg gcagtagagg
     2941 gcatggtgga agagctcatc gccagccatg ctgatgtcaa tgctgtggat gagcttggga
     3001 aatcagcctt acactgggct gcggctgtga acaacgtgga agccactttg gccctgctca
     3061 aaaatggagc caataaggac atgcaggata gcaaggagga gaccccccta ttcctggccg
     3121 cccgcgaggg cagctatgag gctgccaagc tgctgttgga ccactttgcc aaccgtgaga
     3181 tcaccgacca cctggacagg ctgccgcggg acgtagccca ggagagactg caccaggaca
     3241 tcgtgcgctt gctggatcaa cccagtgggc cccgcagccc ccccggtccc cacggcctgg
     3301 ggcctctgct ctgtcctcca ggggccttcc tccctggcct caaagcggca cagtcggggt
     3361 ccaagaagag caggaggccc cccgggaagg cggggctggg gccgcagggg ccccgggggc
     3421 ggggcaagaa gctgacgctg gcctgcccgg gccccctggc tgacagctcg gtcacgctgt
     3481 cgcccgtgga ctcgctggac tccccgcggc ctttcggtgg gccccctgct tcccctggtg
     3541 gcttccccct tgaggggccc tatgcagctg ccactgccac tgcagtgtct ctggcacagc
     3601 ttggtggccc aggccgggcg ggtctagggc gccagccccc tggaggatgt gtactcagcc
     3661 tgggcctgct gaaccctgtg gctgtgcccc tcgattgggc ccggctgccc ccacctgccc
     3721 ctccaggccc ctcgttcctg ctgccactgg cgccgggacc ccagctgctc aacccaggga
     3781 cccccgtctc cccgcaggag cggcccccgc cttacctggc agtcccagga catggcgagg
     3841 agtacccggc ggctggggca cacagcagcc ccccaaaggc ccgcttcctg cgggttccca
     3901 gtgagcaccc ttacctgacc ccatcccccg aatcccctga gcactgggcc agcccctcac
     3961 ctccctccct ctcagactgg tccgaatcca cgcctagccc agccactgcc actggggcca
     4021 tggccaccac cactggggca ctgcctgccc agccacttcc cttgtctgtt cccagctccc
     4081 ttgctcaggc ccagacccag ctggggcccc agccggaagt tacccccaag aggcaagtgt
     4141 tggcctgaga cgctcgtcag ttcttagatc gatctggtag gaattccccc cctaacgtta
     4201 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     4261 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     4321 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     4381 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     4441 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     4501 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     4561 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     4621 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     4681 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     4741 gataatatgg ggtcgaggct gctgctgctg ctgctgctgg gcctgaggct acagctctcc
     4801 ctgggcatca tcccagttga ggaggagaac ccggacttct ggaaccgcga ggcagccgag
     4861 gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa cctcatcatc
     4921 ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct aaaagggcag
     4981 aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc atatgtggct
     5041 ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac agccacggcc
     5101 tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc cgcccgcttt
     5161 aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa
     5221 gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc gccagccggc
     5281 acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc
     5341 cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga cattgacgtg
     5401 atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc tgagtaccca
     5461 gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca ggaatggctg
     5521 gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca ggcttccctg
     5581 gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa atacgagatc
     5641 caccgagact ccacactgga cccctccctg atggagatga cagaggctgc cctgcgcctg
     5701 ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt
     5761 catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga cgacgccatt
     5821 gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac tgccgaccac
     5881 tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt cgggctggcc
     5941 cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa cggtccaggc
     6001 tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg gagccccgag
     6061 tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga ggacgtggcg
     6121 gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca gaccttcata
     6181 gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga cctggcgccc
     6241 cccgccggca ccaccgacgc cgcgcacccg gggcggtccg tggtccccgc gttgcttcct
     6301 ctgctggccg ggaccctgct gctgctggag acggccactg ctccctgagt gtcccgtccc
     6361 tggggctcct gcttccccat cccggagttc tcctgctccc cacctcctgt cgtcctgcct
     6421 ggcctccagc ccgagtcgtc atccccggag tccctataca gaggtcctgc catggaacct
     6481 tcccctcccc gtgcgctctg gggactgagc ccatgacacc aaacctgccc cttggctgct
     6541 ctcggactcc ctaccccaac cccagggact gcaggttgtg ccctgtggct gcctgcaccc
     6601 caggaaagga gggggctcag gccatccagc caccacctac agcccagtgg ggtcgacccg
     6661 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     6721 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     6781 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     6841 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     6901 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6961 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     7021 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     7081 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     7141 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     7201 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     7261 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     7321 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     7381 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7441 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7501 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8281 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9301 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9361 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9421 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9481 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9541 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     9601 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     9661 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cgcgtt
//
