LOCUS       sequence-319262-        7757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses rat PAM3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7757)
  AUTHORS   Milgram SL, Eipper BA, Mains RE
  TITLE     Differential trafficking of soluble and integral membrane secretory 
            granule-associated proteins.
  JOURNAL   J Cell Biol. 1994 Jan;124(1-2):33-41. doi: 10.1083/jcb.124.1.33.
  PUBMED    8294504
REFERENCE   2  (bases 1 to 7757)
  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 Cell 
            Biol."; date: "1994-01"; volume: "124(1-2):33-41. doi"; pages: " 
            10.1083/jcb.124.1.33"
COMMENT     Sequence Label: pCIS-rPAM3
FEATURES             Location/Qualifiers
     source          1..7757
                     /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(3295..3311)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3295..3311)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    3323..3457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3525..3882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3525..3716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3548..3568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3733..3868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3923..4486
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(5211..5230)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(5917..6505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5997..6016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6676..7467,7468..7536))
                     /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     7299..7318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7537..7641)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7709..7727
                     /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 gaggtttaaa gaaactacca gatcattttc caatgaatgc cttggtacca ttggaccagt
     1081 cacccctctt gatgcatcag attttgcgct ggatattcgc atgcctgggg ttacacctaa
     1141 agagtctgac acatacttct gcatgtccat gcgtctgcct gtggatgagg aagccttcgt
     1201 gattgacttc aagcctcgtg ccagcatgga tactgtccac catatgctgc tgtttggatg
     1261 caatatgccc tcgtccactg gaagttactg gttttgtgat gaaggaacct gtacagataa
     1321 agccaatatt ctatatgcct gggcaaggaa tgctcccccc acccggctcc cgaaaggtgt
     1381 tggattcaga gttggaggag aaactggaag caaatacttc gtccttcaag ttcactatgg
     1441 cgatatcagt gcttttcgag ataatcacaa agactgctct ggcgtgtccg tacatctcac
     1501 acgtgtgccc cagcctttaa ttgcgggcat gtaccttatg atgtctgttg acactgtcat
     1561 accaccagga gagaaagtga atgctgacat ttcgtgccaa tacaaaatgt atccaatgca
     1621 tgtgtttgcc tacagagtcc acactcacca tttaggtaag gtggtgagcg gatacagagt
     1681 aagaaacgga cagtggacac tgattggacg ccagaacccc cagctgccac aggctttcta
     1741 ccctgtggaa caccccgttg atgttacttt tggtgatata ctggcagcca gatgtgtgtt
     1801 cactggtgaa gggaggacag aggccaccca tatcggcggc acttctagtg acgaaatgtg
     1861 taacctgtac atcatgtatt acatggaagc caaatatgca ctttccttca tgacctgtac
     1921 aaagaacgtg gctccagata tgttcagaac tatcccagca gaggccaata tcccaattcc
     1981 tgtcaaaccg gacatggtta tgatgcacgg gcatcacaaa gaagcagaaa acaaagaaaa
     2041 gagtgcttta atgcagcagc caaaacaggg agaggaagaa gtattagagc aggatttcca
     2101 tgtggaagaa gaactggact ggcctggagt gtacttgtta ccaggccagg tttctggggt
     2161 ggccctggat tctaagaata acctagtgat tttccacaga ggtgaccatg tttgggatgg
     2221 aaactctttt gacagcaagt ttgtttacca gcaaagaggt cttgggccaa ttgaagaaga
     2281 caccatcctg gtcattgacc caaataatgc tgaaatcctc cagtccagtg gcaagaacct
     2341 gttttattta ccacacggct tgagcataga tacagatgga aattattggg tcacagatgt
     2401 ggctctccac caggtgttca aattggaccc gcatagcaaa gaaggccctc tcttaattct
     2461 gggaaggagc atgcaacctg ggagtgacca aaatcatttc tgccagccca ccgatgtggc
     2521 tgtggagccc agtactggag ctgtcttcgt gtcagacggt tactgtaaca gtcggattgt
     2581 gcagttttca ccaagcggaa agttcgtcac ccagtgggga gaagagtcct ctggaagcag
     2641 tcctaggcca ggccagttca gtgttcctca cagtttggcc cttgtgcctc atttggacca
     2701 gttgtgtgtg gcagacaggg aaaatggccg aatccaatgc ttcaaaactg acaccaaaga
     2761 atttgtgaga gagattaagc acgcatcatt tggaaggaat gtctttgcca tttcatatat
     2821 accaggtttc ctctttgccg taaacgggaa gccttacttt ggagaccaag agcccgtgca
     2881 aggatttgtg atgaactttt ccagtgggga aattatagac gtcttcaagc cagtacgcaa
     2941 gcacttcgac atgcctcatg atattgtggc ttctgaagat gggactgtgt acattggaga
     3001 cgcacacaca aacaccgtgt ggaagttcac cctgactgaa aaaatggagc atcggtcagt
     3061 taaaaaggct ggcattgaag tccaggaaat caaaggaaag ggaagcggcg gcttaaatct
     3121 gggaaatttc tttgcaagtc gaaaaggcta cagcagaaaa gggtttgacc gagtgagcac
     3181 agaggggagt gaccaagaga aagatgagga cgacggaact gagtctgaag aggagtactc
     3241 ggccccgctg cccaagcctg caccttcctc ctgagctcca gccttcgccc gggggatcca
     3301 ctagttctag agcggccgcg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttatcatgt ctggatcccc gggaattcgg cgcagcacca
     3481 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3541 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttat cgcgataaga
     3901 ggattttatc cccgctgcca tcatggttcg accattgaac tgcatcgtcg ccgtgtccca
     3961 aaatatgggg attggcaaga acggagacct accctggcct ccgctcagga acgagttcaa
     4021 gtacttccaa agaatgacca caacctcttc agtggaaggt aaacagaatc tggtgattat
     4081 gggtaggaaa acctggttct ccattcctga gaagaatcga cctttaaagg acagaattaa
     4141 tatagttctc agtagagaac tcaaagaacc accacgagga gctcattttc ttgccaaaag
     4201 tttggatgat gccttaagac ttattgaaca accggaattg gcaagtaaag tagacatggt
     4261 ttggatagtc ggaggcagtt ctgtttacca ggaagccatg aatcaaccag gccacctcag
     4321 actctttgtg acaaggatca tgcaggaatt tgaaagtgac acgtttttcc cagaaattga
     4381 tttggggaaa tataaacttc tcccagaata cccaggcgtc ctctctgagg tccaggagga
     4441 aaaaggcatc aagtataagt ttgaagtcta cgagaagaaa gactaacagg aagatgcttt
     4501 caagttctct gctcccctcc taaagctatg catttttata agaccatggg acttttgctg
     4561 gctttagatc cttcgcggga cgtcctttgt ttacgtcccg tcggcgctga atcccggacg
     4621 acccctcgcg gggccgcttg ggactctctc gtccccttct ccgtctgccg ttccagccga
     4681 ccacggggcg cacctctctt tacgcggtct ccccgtctgt gccttctcat ctgccggtcc
     4741 gtgtgcactt cgcttcacct ctgcacgttg catggagacc accgtgaacg cccatcagat
     4801 cctgcccaag gtcttacata agaggactct tggactccca gcaatgtcaa cgaccgacct
     4861 tgaggcctac ttcaaagact gtgtgtttaa ggactgggag gagctggggg aggagattag
     4921 gttaaaggtc tttgtattag gaggctgtag gcataaattg gtctgcgcac cagcaccatg
     4981 caactttttc acctctgcct aatcatctct tgtacatgtc ccactgttca agcctccaag
     5041 ctgtgccttg ggtggctttg gggcatggac attgaccctt ataaagaatt tggagctact
     5101 gtggagttac tctcgttttt gccttctgac ttctttcctt ccgtcagaga tcctctacgc
     5161 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     5221 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     5281 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     5341 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     5401 gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca acccagtcag
     5461 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     5521 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     5581 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     5641 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     5701 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     5761 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     5821 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gccagcaaaa
     5881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6961 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7261 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7681 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     7741 agaattcgag ctcgccc
//
