LOCUS       sequence-191056-        5772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression with secretive signal and FC tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  AUTHORS   Aricescu AR, Lu W, Jones EY
  TITLE     A time- and cost-efficient system for high-level protein production 
            in mammalian cells.
  JOURNAL   Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50. 
            Epub 2006 Sep 19.
  PUBMED    17001101
REFERENCE   2  (bases 1 to 5772)
  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: "Acta 
            Crystallogr D Biol Crystallogr."; date: "2006-10"; volume: "62(Pt 
            10)"; pages: "1243-50. Epub 2006 Sep 19"
COMMENT     Sequence Label: pHL-FcHis
FEATURES             Location/Qualifiers
     source          1..5772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2289..2312
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3003..3020
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3126..3145)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3191..3246
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3245..3264)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3607..3623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3607..3623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3620..3642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3631..3647
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3655..3661,3662..3679,3680..3685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3700..3721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3850..3867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4021..4609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4780..5571,5572..5640))
                     /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     5403..5422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5641..5745)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc
     1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggtt
     1861 cgatagtttt agagcctatc tattggaatt cctcgaactc caaatttcta cctggacaag
     1921 gactggtact atacccacag ataggagaca aattggatat tatttgcccc aaagtggact
     1981 ctaaaactgt tggccagtat gaatattata aagtttatat ggttgataaa gaccaagcag
     2041 acagatgcac tattaagaag gaaaataccc ctctcctcaa ctgtgccaaa ccggaccaag
     2101 atatcaaatt caccatcaag tttcaagaat tcagccctaa cctctggggt ctagaatttc
     2161 agaagaacaa agattattac attatatcta catcaaatgg gtctttggag ggcctggata
     2221 accaggaggg aggggtgtgc cagacaagag ccatgaagat cctcatgaaa gttggacaag
     2281 atggtaccct ggaggtgctg ttccagggcc ccaaatcttg tgacaaaact cacacatgcc
     2341 caccgtgccc agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac
     2401 ccaaggacac cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga
     2461 gccacgaaga ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg
     2521 ccaagacaaa gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca
     2581 ccgtcctgca ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag
     2641 ccctcccagc ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac
     2701 aggtgtacac cctgccccca tcccgggatg agctgaccaa gaaccaggtc agcctgacct
     2761 gcctggtcaa aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc
     2821 cggagaacaa ctacaaggcc acgcctcccg tgctggactc cgacggctcc ttcttcctct
     2881 acagcaagct caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg
     2941 tgatgcatga ggctctgcac aaccactaca cgcagaagag cctctccctg tctccgggta
     3001 aacatcacca tcatcaccat tgactcgaga ctagtatcgc gataattcac tcctcaggtg
     3061 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     3121 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     3181 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     3241 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     3301 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     3361 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     3421 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     3481 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     3541 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     3601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga
     3781 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4441 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5761 agtgccacct gg
//
