LOCUS       sequence-246463-        8431 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses N-terminal Flag-tagged human CNP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8431)
  AUTHORS   Unlu I, Lu Y, Wang X
  TITLE     The cyclic phosphodiesterase CNP and RNA cyclase RtcA fine-tune 
            noncanonical XBP1 splicing during ER stress.
  JOURNAL   J Biol Chem. 2018 Dec 14;293(50):19365-19376. doi: 
            10.1074/jbc.RA118.004872. Epub 2018 Oct 24.
  PUBMED    30355738
REFERENCE   2  (bases 1 to 8431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.RA118.004872"; journalName: "J Biol Chem"; date: 
            "2018-12-14- 14"; volume: "293"; issue: "50"; pages: "19365-19376"
COMMENT     Sequence Label: pB-bsd-Flag-CNP
FEATURES             Location/Qualifiers
     source          1..8431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..1143
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1067..1089
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1151..1170
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     polyA_signal    2589..2813
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3186..3248
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3314..3333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3315..3333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3340..3357)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3340..3356)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3349..3371)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3498..3953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3585..3604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3795..3816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3979..4083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4084..4152,4153..4944)
                     /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(4302..4321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5115..5703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5604..5623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5857..5874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5991..6012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6027..6032,6033..6050,6051..6057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6065..6081
                     /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)."
     primer_bind     6070..6092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6089..6105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6089..6105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6126..6144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   6293..6327
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    6728..6952
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(6980..7378)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MGMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7375..7384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7398..7897)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7446..7465)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7839..7862
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        7917..8296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8298..8431,1..126)
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      301 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      361 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      421 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
      481 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
      541 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
      601 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
      661 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
      721 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
      781 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
      841 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      901 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      961 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1021 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1081 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1141 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat
     1201 cagcttggca aagcattcat ggactacaaa gaccatgacg gtgattataa agatcatgac
     1261 atcgattaca aggatgacga tgaccggatc cgaattctaa tgaacactag ctttacccgc
     1321 aaaagccaca cattcctgcc caagctcttc ttcaggaaaa tgtcatcctc aggagcaaag
     1381 gagaagccag agctgcagtt ccctttcctt caagatgagg acacggtggc taccctccac
     1441 gagtgcaaga cgctattcat cctgcgcggc ctgccgggca gcggcaagtc cacactggct
     1501 cggctcatcc tggagaagta ccacgacggc accaagatgg tgtccgctga tgcttacaag
     1561 atcatccctg gctctcgggc agacttctcc gaggcgtaca agcgtctaga cgaggacctg
     1621 gctggctact gccgccggga catcagggtt cttgtgcttg atgataccaa ccacgagcgg
     1681 gagcggctgg atcagctttt tgaaatggca gaccagtatc agtaccaggt ggtgctggtg
     1741 gagcccaaga cagcgtggcg actagactgt gcccagctca aggagaagaa ccaatggcag
     1801 ctgtcggccg atgacctgaa gaagctgaag cccgggctgg agaaggactt tctgccactc
     1861 tactttggct ggttcctgac caaaaagagt tctgagaccc tccgaaaagc tggccaggtc
     1921 tttctggagg agctggggaa tcacaaggcc ttcaagaaag agcttcgaca ctttatttct
     1981 ggagatgaac ccaaggagaa gcttgagctg gtcagctact ttggaaagag acctccaggt
     2041 gtgctgcact gtacaaccaa attctgtgac tacgggaagg ccgctggggc agaagaatac
     2101 gcccagcagg aggtggtgaa gagatcgtat ggcaaggcct tcaaactgtc catctctgct
     2161 ctctttgtga cacccaagac agctggggcc caggtggtgc tgaccgatca ggagctgcag
     2221 ttgtggccaa gtgatctgga caagccatct gcctccgagg gcctgccccc agggagccga
     2281 gctcacgtca ccctaggctg tgcggccgac gtgcagccag tgcagacggg tcttgacctc
     2341 ttagatattt tacaacaggt gaaggggggc agccaaggtg aggcggtggg tgagctcccc
     2401 cggggcaagc tctattccct gggcaaaggg cgatggatgc tgagcctgac taagaagatg
     2461 gaggtcaagg ccatcttcac ggggtactat gggaagggca aacctgtgcc catacatggc
     2521 agccggaagg ggggtgccat gcagatctgc accatcatct gagctcgagg gccgctgatc
     2581 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2641 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2701 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2761 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2821 ggcggaaaga accagctggg gcttaattaa cgagagcata atattgatat gtgccaaagt
     2881 tgtttctgac tgactaataa gtataatttg tttctattat gtataggtta agctaattac
     2941 ttattttata atacaacatg actgttttta aagtacaaaa taagtttatt tttgtaaaag
     3001 agagaatgtt taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt
     3061 aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa
     3121 atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata
     3181 aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct
     3241 ttctagggtt aaataatagt ttctaatttt tttattattc agcctgctgt cgtgaatacc
     3301 gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     3361 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3421 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3481 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3541 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3601 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3661 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3721 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3781 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3841 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3901 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6121 ctcgaaatta accctcacta aagggaacaa aagctggtac ctcgcgcgac ttggtttgcc
     6181 attctttagc gcgcgtcgcg tcacacagct tggccacaat gtggtttttg tcaaacgaag
     6241 attctatgac gtgtttaaag tttaggtcga gtaaagcgca aatctttttt aaccctagaa
     6301 agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata
     6361 gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg
     6421 cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa
     6481 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
     6541 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
     6601 cgtgactaat atataataaa atgggtagtt ctttagacga tgagcatatc ctctctgctc
     6661 ttctgcaaag cgatgacgag cttgttggct agcgtcgagc cccagctggt tctttccgcc
     6721 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     6781 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     6841 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     6901 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     6961 gatcagcgag ctctagagct tagccctccc acacataacc agagggcagc aattcacgaa
     7021 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     7081 gatcgagaag cacctgtcga caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     7141 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     7201 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     7261 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     7321 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc atccccatgg
     7381 tggcgttggc tgcaggtcga aaggcccgga gatgaggaag aggagaacag cgcggcagac
     7441 gtgcgctttt gaagcgtgca gaatgccggg cctccggagg accttcgggc gcccgccccg
     7501 cccctgagcc cgcccctgag cccgcccccg gacccacccc ttcccagcct ctgagcccag
     7561 aaagcgaagg agcaaagctg ctattggccg ctgccccaaa ggcctacccg cttccattgc
     7621 tcagcggtgc tgtccatctg cacgagacta gtgagacgtg ctacttccat ttgtcacgtc
     7681 ctgcacgacg cgagctgcgg ggcggggggg aacttcctga ctaggggagg agtagaaggt
     7741 ggcgcgaagg ggccaccaaa gaacggagcc ggttggcgcc taccggtgga tgtggaatgt
     7801 gtgcgaggcc agaggccact tgtgtagcgc caagtgccca gcggggctgc taaagcgcat
     7861 gctccagact gccttgggaa aagcgcctcc cctacccggt agaattaatt cgattcgaca
     7921 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     7981 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8041 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8101 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8161 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8221 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8281 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     8341 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     8401 ggcggggggg gggggggggg ggcggggcga g
//
