LOCUS       sequence-299336-        6465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the catalytic core of rat 
            peptidyl-hydroxyglycine-alpha-amidating lyase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Kolhekar AS, Bell J, Shiozaki EN, Jin L, Keutmann HT, Hand TA, Mains
            RE, Eipper BA
  TITLE     Essential features of the catalytic core of 
            peptidyl-alpha-hydroxyglycine alpha-amidating lyase.
  JOURNAL   Biochemistry. 2002 Oct 15;41(41):12384-94. doi: 10.1021/bi0260280.
  PUBMED    12369828
REFERENCE   2  (bases 1 to 6465)
  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"; doi: "10"; journalName: 
            "Biochemistry"; date: "2002-10-15- 15"; volume: "41"; issue: "41"; 
            pages: "12384-94"
COMMENT     Sequence Label: pCIS-rPALcc
FEATURES             Location/Qualifiers
     source          1..6465
                     /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"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      914..923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2003..2019)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2019)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    2031..2165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2068..2087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2122..2141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2233..2590
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2233..2424
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2256..2276)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2441..2576
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2631..3194
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(3919..3938)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(4625..5213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4705..4724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5384..6175,6176..6244))
                     /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     6007..6026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6245..6349)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6417..6435
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcttatgcg acggatcccg
      781 taataagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg acaatgacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagc tcggttctat
      901 cgataagctt gccgccacca tggccggacg cgcccgcagc ggtctgctac tgctgctgct
      961 ggggctgctc gccctgcaga gcagctgcct ggccttcaga agcccacttt ctgtctttaa
     1021 gaggcatatg gatttccatg tggaagaaga actggactgg cctggagtgt acttgttacc
     1081 aggccaggtt tctggggtgg ccctggattc taagaataac ctagtgattt tccacagagg
     1141 tgaccatgtt tgggatggaa actcttttga cagcaagttt gtttaccagc aaagaggtct
     1201 tgggccaatt gaagaagaca ccatcctggt cattgaccca aataatgctg aaatcctcca
     1261 gtccagtggc aagaacctgt tttatttacc acacggcttg agcatagata cagatggaaa
     1321 ttattgggtc acagatgtgg ctctccacca ggtgttcaaa ttggacccgc atagcaaaga
     1381 aggccctctc ttaattctgg gaaggagcat gcaacctggg agtgaccaaa atcatttctg
     1441 ccagcccacc gatgtggctg tggagcccag tactggagct gtcttcgtgt cagacggtta
     1501 ctgtaacagt cggattgtgc agttttcacc aagcggaaag ttcgtcaccc agtggggaga
     1561 agagtcctct ggaagcagtc ctaggccagg ccagttcagt gttcctcaca gtttggccct
     1621 tgtgcctcat ttggaccagt tgtgtgtggc agacagggaa aatggccgaa tccaatgctt
     1681 caaaactgac accaaagaat ttgtgagaga gattaagcac gcatcatttg gaaggaatgt
     1741 ctttgccatt tcatatatac caggtttcct ctttgccgta aacgggaagc cttactttgg
     1801 agaccaagag cccgtgcaag gatttgtgat gaactttgcc agtggggaaa ttatagacgt
     1861 cttcaagcca gtacgcaagc acttcgacat gcctcatgat attgtggctt ctgaagatgg
     1921 gactgtgtac attggagacg cacacacaaa caccgtgtgg aagttcaccc tgactgaaaa
     1981 aatggagcat cggtcagttt aggatccact agttctagag cggccgcgtt aacttgttta
     2041 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2101 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2161 ggatccccgg gaattcggcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     2221 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2281 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2341 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2401 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2461 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2521 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2581 cttttgcaaa aagcttatcg cgataagagg attttatccc cgctgccatc atggttcgac
     2641 cattgaactg catcgtcgcc gtgtcccaaa atatggggat tggcaagaac ggagacctac
     2701 cctggcctcc gctcaggaac gagttcaagt acttccaaag aatgaccaca acctcttcag
     2761 tggaaggtaa acagaatctg gtgattatgg gtaggaaaac ctggttctcc attcctgaga
     2821 agaatcgacc tttaaaggac agaattaata tagttctcag tagagaactc aaagaaccac
     2881 cacgaggagc tcattttctt gccaaaagtt tggatgatgc cttaagactt attgaacaac
     2941 cggaattggc aagtaaagta gacatggttt ggatagtcgg aggcagttct gtttaccagg
     3001 aagccatgaa tcaaccaggc cacctcagac tctttgtgac aaggatcatg caggaatttg
     3061 aaagtgacac gtttttccca gaaattgatt tggggaaata taaacttctc ccagaatacc
     3121 caggcgtcct ctctgaggtc caggaggaaa aaggcatcaa gtataagttt gaagtctacg
     3181 agaagaaaga ctaacaggaa gatgctttca agttctctgc tcccctccta aagctatgca
     3241 tttttataag accatgggac ttttgctggc tttagatcct tcgcgggacg tcctttgttt
     3301 acgtcccgtc ggcgctgaat cccggacgac ccctcgcggg gccgcttggg actctctcgt
     3361 ccccttctcc gtctgccgtt ccagccgacc acggggcgca cctctcttta cgcggtctcc
     3421 ccgtctgtgc cttctcatct gccggtccgt gtgcacttcg cttcacctct gcacgttgca
     3481 tggagaccac cgtgaacgcc catcagatcc tgcccaaggt cttacataag aggactcttg
     3541 gactcccagc aatgtcaacg accgaccttg aggcctactt caaagactgt gtgtttaagg
     3601 actgggagga gctgggggag gagattaggt taaaggtctt tgtattagga ggctgtaggc
     3661 ataaattggt ctgcgcacca gcaccatgca actttttcac ctctgcctaa tcatctcttg
     3721 tacatgtccc actgttcaag cctccaagct gtgccttggg tggctttggg gcatggacat
     3781 tgacccttat aaagaatttg gagctactgt ggagttactc tcgtttttgc cttctgactt
     3841 ctttccttcc gtcagagatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     3901 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     3961 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     4021 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     4081 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaccga
     4141 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     4201 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     4261 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     4321 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     4381 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
     4441 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     4501 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     4561 atcagggaca gcttcaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5041 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5281 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5341 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5401 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5461 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5521 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5581 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5641 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     5701 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5761 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5821 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5881 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5941 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6001 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6061 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6121 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6181 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6241 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6361 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6421 ggcgtatcac gaggcccttt cgtcttcaag aattcgagct cgccc
//
