LOCUS       sequence-8222-ex        6505 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 6505)
  AUTHORS   Bao ZZ, Cepko CL
  TITLE     The expression and function of Notch pathway genes in the developing
            rat eye.
  JOURNAL   J Neurosci. 1997 Feb 15. 17(4):1425-34.
  PUBMED    9006984
REFERENCE   2  (bases 1 to 6505)
  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.
            1997 Feb 15. 17(4):1425-34."
COMMENT     Sequence Label: pLIA (CC#1)
FEATURES             Location/Qualifiers
     source          1..6505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1008..1799,1800..1868))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1631..1650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1869..1973)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2539..2738
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2939..3355
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3285..3307
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3323..3339
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3417..3983
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3434..3985
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3578..3595)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3805..3824
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4007..5506
                     /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="LLLLLLLGLRLQLSLGIILVEEENPDFWNREAAEALGAAKKLQPA
                     QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
                     VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
                     TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
                     MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLGERQGARYVWNRTELMQASLDPSVTHL
                     MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRHPRGFFLFVEGGRIDHGHHESRA
                     YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSFIGLAAGKAR
                     DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
                     GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     LTR             5911..6504
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttgcggcc
      961 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1141 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1321 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1621 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1861 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1981 ccccgaaaag tgccacctga cgtgctgcag aatatgggcc aaacaggata tctgtggtaa
     2041 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     2101 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     2161 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgcccccaag
     2221 aacctgaaat gaccctggcc ttatttgaac taaccaatca gttcgtttct cgtttctgtt
     2281 ggggcgcttc tgttcccgga gctcaataaa agagcccaca acccttcatt gggggcgcca
     2341 gtcttccgat tgactgagtc gcccggtacc gtgtatccaa taaaccttct tgcagttgca
     2401 tcagacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg acaacccgtc
     2461 agcggggtct ttcatttggg ggctcgtccg ggatcgggag acccttgccc agggaccacg
     2521 gacccaccac cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt
     2581 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
     2641 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
     2701 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
     2761 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     2821 gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg
     2881 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     2941 ccagactgtt accaatccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     3001 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     3061 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     3121 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     3181 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     3241 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     3301 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggccg
     3361 gatccgcggc cgcataggcc actagtggat cccagtgtgg tggtacgtag gaattctccc
     3421 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     3481 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     3541 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     3601 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     3661 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     3721 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     3781 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     3841 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     3901 acgtgtgttt agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     3961 tcctttgaaa aacacgatga taatatggcc aaatatgggg tcgaggctgc tgctgctgct
     4021 gctgctgggc ctgaggctac agctctccct gggcatcatc ctagttgagg aggagaaccc
     4081 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     4141 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     4201 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     4261 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     4321 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     4381 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     4441 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     4501 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     4561 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     4621 gctcatctcc aacatggaca ttgatgtgat cctaggtgga ggccgaaagt acatgtttcg
     4681 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     4741 cgggaagaat ctggtgcagg aatggctcgg cgaacgccag ggtgcccggt acgtgtggaa
     4801 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     4861 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     4921 ggagatgaca gaggctgccc tgcgcctgct gagcagacac ccccgcggct tcttcctctt
     4981 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     5041 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     5101 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     5161 gcgagggagc tccttcatcg ggctggccgc tggcaaggcc cgggacagga aggcctacac
     5221 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     5281 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     5341 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     5401 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     5461 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     5521 gcggtccgtg gtccccgcgt tgcttcctct gctggccggg accctgctgc tgctggagac
     5581 ggccactgct ccctgagtgt cccgtccctg gggctcctgc ttccccatcc cggagttctc
     5641 ctgctcccca cctcctgtcg tcctgcctgg cctccagccc gagtcgtcat ccccggagtc
     5701 cctatacaga ggtcctgcca tggaaccttc ccctccccgt gcgctctggg gactgagccc
     5761 atgacaccaa acctgcccct tggctgctct cggactccct accccaaccc cagggactgc
     5821 aggttgtgcc ctgtggctgc ctgcacccca ggaaaggagg gggctcaggc catccagcca
     5881 ccacctacag cccagtgggt acggtcgacc aatgaaagac cccacctgta ggtttggcaa
     5941 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     6001 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     6061 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     6121 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     6181 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     6241 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     6301 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     6361 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     6421 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     6481 acccgtcagc gggggtcttt cattt
//
