LOCUS       sequence-299344-        8428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses catalytically inactive rat PAM-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8428)
  AUTHORS   Eipper BA, Quon AS, Mains RE, Boswell JS, Blackburn NJ
  TITLE     The catalytic core of peptidylglycine alpha-hydroxylating 
            monooxygenase: investigation by site-directed mutagenesis, Cu X-ray 
            absorption spectroscopy, and electron paramagnetic resonance.
  JOURNAL   Biochemistry. 1995 Mar 7;34(9):2857-65. doi: 10.1021/bi00009a016.
  PUBMED    7893699
REFERENCE   2  (bases 1 to 8428)
  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: "1995-03-7- 7"; volume: "34"; issue: "9"; 
            pages: "2857-65"
COMMENT     Sequence Label: pCI.neo-rPAM1-M314I
FEATURES             Location/Qualifiers
     source          1..8428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      965..974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3922..3938)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3922..3938)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    3995..4116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4006..4025)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4060..4079
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4302..4757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4389..4408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4599..4620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4874..5231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4874..5065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4897..4917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5082..5217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5282..6076
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5336..5355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5946..5965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6140..6188
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6229..6248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6348..6370
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6408..6426)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6494..6598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6599..6667,6668..7459)
                     /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(6817..6836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7630..8218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8119..8138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt 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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct acgataagct
      961 tgccgccacc atggccggac gcgcccgcag cggtctgcta ctgctgctgc tggggctgct
     1021 cgccctgcag agcagctgcc tggccttcag aagcccactt tctgtcttta agaggtttaa
     1081 agaaactacc agatcatttt ccaatgaatg ccttggtacc attggaccag tcacccctct
     1141 tgatgcatca gattttgcgc tggatattcg catgcctggg gttacaccta aagagtctga
     1201 cacatacttc tgcatgtcca tgcgtctgcc tgtggatgag gaagccttcg tgattgactt
     1261 caagcctcgt gccagcatgg atactgtcca ccatatgctg ctgtttggat gcaatatgcc
     1321 ctcgtccact ggaagttact ggttttgtga tgaaggaacc tgtacagata aagccaatat
     1381 tctatatgcc tgggcaagga atgctccccc cacccggctc ccgaaaggtg ttggattcag
     1441 agttggagga gaaactggaa gcaaatactt cgtccttcaa gttcactatg gcgatatcag
     1501 tgcttttcga gataatcaca aagactgctc tggcgtgtcc gtacatctca cacgtgtgcc
     1561 ccagccttta attgcgggca tgtaccttat gatgtctgtt gacactgtca taccaccagg
     1621 agagaaagta gtgaatgctg acatttcgtg ccaatacaaa atgtatccaa tgcatgtgtt
     1681 tgcctacaga gtccacactc accatttagg taaggtggtg agcggataca gagtaagaaa
     1741 cggacagtgg acactgattg gacgccagaa cccccagctg ccacaggctt tctaccctgt
     1801 ggaacacccc gttgatgtta cttttggtga tatactggca gccagatgtg tgttcactgg
     1861 tgaagggagg acagaggcca cccatatcgg cggcacttct agtgacgaaa tatgtaacct
     1921 gtacatcatg tattacatgg aagccaaata tgcactttcc ttcatgacct gtacaaagaa
     1981 cgtggctcca gatatgttca gaactatccc agcagaggcc aatatcccaa ttcctgtcaa
     2041 accggacatg gttatgatgc acgggcatca caaagaagca gaaaacaaag aaaagagtgc
     2101 tttaatgcag cagccaaaac agggagagga agaagtatta gagcagggtg atttctattc
     2161 actgctttcc aagctgctag gagaaaggga agatgttcat gtgcacaagt ataatcctac
     2221 agaaaagaca gaatctgggt cagacctggt agctgagatt gcaaacgtgg tccagaaaaa
     2281 ggaccttggt cggtctgacg ccagagaagg tgcagagcat gaggaatggg gtaatgctat
     2341 cctagtcaga gacaggatcc acagattcca ccagctagag tcaactctga ggccagctga
     2401 gagcagagct ttctcgttcc agcagcctgg cgaaggccct tgggaaccag aaccctcagg
     2461 agatttccat gtggaagaag aactggactg gcctggagtg tacttgttac caggccaggt
     2521 ttctggggtg gccctggatt ctaagaataa cctagtgatt ttccacagag gtgaccatgt
     2581 ttgggatgga aactcttttg acagcaagtt tgtttaccag caaagaggtc ttgggccaat
     2641 tgaagaagac accatcctgg tcattgaccc aaataatgct gaaatcctcc agtccagtgg
     2701 caagaacctg ttttatttac cacacggctt gagcatagat acagatggaa attattgggt
     2761 cacagatgtg gctctccacc aggtgttcaa attggacccg catagcaaag aaggccctct
     2821 cttaattctg ggaaggagca tgcaacctgg gagtgaccaa aatcatttct gccagcccac
     2881 cgatgtggct gtggagccca gtactggagc tgtcttcgtg tcagacggtt actgtaacag
     2941 tcggattgtg cagttttcac caagcggaaa gttcgtcacc cagtggggag aagagtcctc
     3001 tggaagcagt cctaggccag gccagttcag tgttcctcac agtttggccc ttgtgcctca
     3061 tttggaccag ttgtgtgtgg cagacaggga aaatggccga atccaatgct tcaaaactga
     3121 caccaaagaa tttgtgagag agattaagca cgcatcattt ggaaggaatg tctttgccat
     3181 ttcatatata ccaggtttcc tctttgccgt aaacgggaag ccttactttg gagaccaaga
     3241 gcccgtgcaa ggatttgtga tgaacttttc cagtggggaa attatagacg tcttcaagcc
     3301 agtacgcaag cacttcgaca tgcctcatga tattgtggct tctgaagatg ggactgtgta
     3361 cattggagac gcacacacaa acaccgtgtg gaagttcacc ctgactgaaa aaatggagca
     3421 tcggtcagtt aaaaaggctg gcattgaagt ccaggaaatc aaagaagccg aggcagttgt
     3481 tgaacccaaa gtggagaaca aacccacctc ctcagaattg cagaagatgc aagagaaaca
     3541 gaaactgagc acagagcccg gctcgggagt gtccgtggtt ctcattacaa cccttctggt
     3601 tattcctgtg ctggtcctgc tggccattgt catgtttatt cggtggaaaa aatcaagggc
     3661 ctttggagat catgaccgca agctcgagtc aagttctgga agagtcctgg gaagattccg
     3721 aggaaaggga agcggcggct taaatctggg aaatttcttt gcaagtcgaa aaggctacag
     3781 cagaaaaggg tttgaccgag tgagcacaga ggggagtgac caagagaaag atgaggacga
     3841 cggaactgag tctgaagagg agtactcggc cccgctgccc aagcctgcac cttcctcctg
     3901 agctccagcc ttcgcccggg ggatccacta gttctagagt cgacccgggc ggccgcttcc
     3961 ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa
     4021 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4081 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4141 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4201 tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg
     4261 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     4321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4741 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4801 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     4861 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4921 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4981 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5041 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5101 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5161 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5221 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac
     5281 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5341 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5401 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5461 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5521 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5581 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5641 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5701 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5761 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5821 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5881 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5941 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6001 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6061 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6121 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     6181 tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc
     6241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     6301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     6421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7741 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc
     8281 tcgacagatc ttcaatattg gccattagcc atattattca ttggttatat agcataaatc
     8341 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     8401 ggctcatgtc caatatgacc gccatgtt
//
