LOCUS       sequence-501612-        6235 bp    DNA     circular SYN 03-MAR-2026
DEFINITION  Expresses a truncated parathyroid hormone caged by chemically 
            activated protein domains in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  TITLE     A chemogenetic approach for temporal and cell-specific activation of
            endogenous GPCRs in vivo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6235)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 3, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PTH-CapC
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     560..579
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        560..578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             778..1098
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1105..1131
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1132..1158
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1906..2415
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             2428..2484
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      2509..2518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2515..3225
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2660..2678)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2928..2947
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3256..3273)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3262..3469
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3498..3638
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3498..3627
                     /label=AAV2 ITR
     rep_origin      3713..4168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3800..3819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4010..4031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4185..4204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4304..4326
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4364..4382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4450..4554
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4555..4623,4624..5415)
                     /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(4773..4792)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5586..6174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6075..6094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      541 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct tgccaccatg
      601 aagaccatca tcgccctgag ctacatcttc tgcctggtgt tcgccagcgt gagcgaaatt
      661 cagctgatgc ataacctggg caaacatctg aacagcatgg aacgcgtgga atggctgcct
      721 aatttgtttg ggtatgtgtc aggcggttct ggtactggat ctggttctgg aggttctgga
      781 gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc
      841 tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aagttgattc ctcccgggac
      901 agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa
      961 ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc
     1021 tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg
     1081 gagcttctaa aactggaaag atcttaccca tacgatgtgc cagattacgc ctatccctat
     1141 gacgtccctg attacgctct cgacttccag aaggcctcca gcatagtcta taagaaagag
     1201 ggggaacagg tggagttctc cttcccactc gcctttacag ttgaaaagct gacgggcagt
     1261 ggcgagctgt ggtggcaggc ggagagggct tcctcctcca agtcttggat cacctttgac
     1321 ctgaagaaca aggaagtgtc tgtaaaacgg gttacccagg accctaagct ccagatgggc
     1381 aagaagctcc cgctccacct caccctgccc caggccttgc ctcagtatgc tggctctgga
     1441 aacctcaccc tggcccttga agcgaaaaca ggaaagttgc atcaggaagt gaacctggtg
     1501 gtgatgagag ccactcagct ccagaaaaat ttgacctgtg aggtgtgggg acccacctcc
     1561 cctaagctga tgctgagctt gaaactggag aacaaggagg caaaggtctc gaagcgggag
     1621 aaggcggtgt gggtgctgaa ccctgaggcg gggatgtggc agtgtctgct gagtgactcg
     1681 ggacaggtcc tgctggaatc caacatcaag gttctgccca catggtccac cccggtgcag
     1741 ccaatggccc tgattgtgct ggggggcgtc gccggcctcc tgcttttcat tgggctaggc
     1801 atcttcttct gtgtcaggtg ccggcaccga aggcgcaagg gctcgggctc gacctcgggc
     1861 tcgggcagcg gtggctctag aggttctggt ggcagctctg gaggtatgaa tggagctata
     1921 ggaggtgacc ttttgctcaa ttttcctgac atgtcggtcc tagagcgcca aagggctcac
     1981 ctcaagtacc tcaatcccac ctttgattct cctctcgccg gcttctttgc cgattcttca
     2041 atgattaccg gcggcgagat ggacagctat ctttcgactg ccggtttgaa tcttccgatg
     2101 atgtacggtg agacgacggt ggaaggtgat tcaagactct caatttcgcc ggaaacgacg
     2161 cttgggactg gaaatttcaa ggcagcgaag tttgatacag agactaagga ttgtaatgag
     2221 gcggcgaaga agatgacgat gaacagagat gacctagtag aagaaggaga agaagagaag
     2281 tcgaaaataa cagagcaaaa caatgggagc acaaaaagca tcaagaagat gaaacacaaa
     2341 gccaagaaag aagagaacaa tttctctaat gattcatcta aagtgacgaa ggaattggag
     2401 aaaacggatt atattcatgg aagcggagct actaacttca gcctgctgaa gcaggctgga
     2461 gacgtggagg agaaccctgg acctaccggt gctcgggatc ctccggtcgc caccatggtg
     2521 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     2581 atggagggct ccgtgaacgg ccacgagttc gagattgaag gtgagggaga gggccgcccc
     2641 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     2701 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     2761 gacatccccg actacttgaa gctgtccttc cccgagggat tcaagtggga gcgcgtgatg
     2821 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     2881 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     2941 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     3001 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     3061 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     3121 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     3181 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt cctcgagcta
     3241 gagctcgctg accagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3301 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3361 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3421 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag gtaaccacgt
     3481 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3541 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3601 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3661 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3721 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3781 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3901 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3961 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4021 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4081 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4141 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4201 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4261 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4321 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4381 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4441 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4501 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4561 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4621 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4681 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4741 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4801 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4861 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4921 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4981 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5041 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5101 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5161 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5221 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
     5281 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5341 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5401 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5461 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5521 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5581 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5641 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5701 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5761 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5821 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5881 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5941 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6001 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6061 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6121 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6181 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
