LOCUS       pcDNA5TO_HsTPC2-        9205 bp    DNA     circular SYN 13-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9205)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     misc_feature    1005..3260
                     /label=HsTPC2
                     /note="color: #a6acb3"
     misc_feature    3270..4595
                     /label=RCaMP181
                     /note="color: #ff0000"
     CDS             3276..3293
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             3297..3329
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /translation="MASMTGGQQMG"
     CDS             join(3333..3341,3342..3356)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /note="This feature has 2 segments:
                        1: 3333 .. 3341 / #cc99b2
                        2: 3342 .. 3356 / #cc99b2 / enterokinase site
                     Cleavage site after base 3356"
                     /translation="DLYDDDDK"
     polyA_signal    4633..4857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4903..5331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5345..5674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5525..5660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             5723..6748
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #ccffcc"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6878..6999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(7048..7064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    7072..7088
                     /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)."
                     /note="color: #31849b"
     promoter        complement(join(7096..7102,7103..7120,7121..7126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 7096 .. 7102 / #ffffff / -10
                        2: 7103 .. 7120 / #ffffff
                        3: 7121 .. 7126 / #ffffff / -35"
     protein_bind    7141..7162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(7450..8038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(join(8209..9000,9001..9069))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 8209 .. 9000 / #ccffcc
                        2: 9001 .. 9069 / #ccffcc / signal sequence
                     Cleavage site after base 9000"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9070..9174)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc caccatggcg gaaccccagg
     1021 cggagtcgga gcccctgctg ggcggggccc gcggcggtgg cggcgactgg ccggcggggc
     1081 tgaccactta ccgcagcatc caagtcggcc ctggtgccgc ggccaggtgg gacctctgca
     1141 ttgatcaggc tgtggtcttc atcgaagatg ctattcagta ccgctccatc aaccaccggg
     1201 tggatgccag ctcgatgtgg ctttaccgac ggtattactc gaacgtatgc caacggactt
     1261 tgagcttcac catcttcttg atcctgtttt tggcttttat cgagacccca tcctcactca
     1321 ccagcacggc ggacgtgcgc taccgcgctg ccccctggga gccgccctgc ggcctgaccg
     1381 agagtgtcga ggtgctctgc ctgctggtct ttgcggccga cctctctgtg aagggttacc
     1441 tgttcgggtg ggcccatttc cagaaaaacc tttggctgct gggctacctc gtggtgctgg
     1501 tggtgtctct ggtggactgg accgtgtccc tgagtctcgt gtgtcatgag cccctgcgga
     1561 tccgccggct tctccgtccc ttcttcctgc tgcagaactc ctctatgatg aagaagacct
     1621 tgaaatgcat ccgctggtcg ctgccggaaa tggccagcgt cgggctgctg ctggccatcc
     1681 acctgtgcct cttcaccatg ttcggaatgc tgctgttcgc tggtgggaag caggatgatg
     1741 ggcaggacag ggagaggctg acctacttcc agaacctgcc tgagtctctg acttccctcc
     1801 tggtgctgct gaccacggcc aacaaccccg atgtgatgat tcctgcgtat tccaagaacc
     1861 gggcctatgc catcttcttc atagtcttca ctgtgatagg aagcctgttt ctgatgaacc
     1921 tgctgacagc catcatctac agtcagttcc ggggctacct gatgaaatct ctccagacct
     1981 cgctgtttcg gaggcggctg ggaacccggg ctgcctttga agtcctatcc tccatggtgg
     2041 gggagggagg agccttccct caggcagttg gggtgaagcc ccagaacttg ctgcaggtgc
     2101 ttcagaaggt ccagctggac agctcccaca aacaggccat gatggagaag gtgcgttcct
     2161 acggcagtgt tctgctgtca gctgaggagt ttcagaagct cttcaacgag cttgacagaa
     2221 gtgtggttaa agagcacccg ccgaggcccg agtaccagtc tccgtttctg cagagcgccc
     2281 agttcctctt cggccactac tactttgact acctggggaa cctcatcgcc ctggcaaacc
     2341 tggtgtccat ttgcgtgttc ctggtgctgg atgcagatgt gctgcctgct gagcgtgatg
     2401 acttcatcct ggggattctc aactgcgtct tcattgtgta ctacctgttg gagatgctgc
     2461 tcaaggtctt tgccctgggc ctgcgagggt acctgtccta ccccagcaac gtgtttgacg
     2521 ggctcctcac cgttgtcctg ctggttttgg agatctcaac tctggctgtg taccgattgc
     2581 cacacccagg ctggaggccg gagatggtgg gcctgctgtc gctgtgggac atgacccgca
     2641 tgctgaacat gctcatcgtg ttccgcttcc tgcgtatcat ccccagcatg aagccgatgg
     2701 ccgtggtggc cagtaccgtc ctgggcctgg tgcagaacat gcgtgctttt ggcgggatcc
     2761 tggtggtggt ctactacgta tttgccatca ttgggatcaa cttgtttaga ggcgtcattg
     2821 tggctcttcc tggaaacagc agcctggccc ctgccaatgg ctcggcgccc tgtgggagct
     2881 tcgagcagct ggagtactgg gccaacaact tcgatgactt tgcggctgcc ctggtcactc
     2941 tgtggaactt gatggtggtg aacaactggc aggtgtttct ggatgcatat cggcgctact
     3001 caggcccgtg gtccaagatc tattttgtat tgtggtggct ggtgtcgtct gtcatctggg
     3061 tcaacctgtt tctggccctg attctggaga acttccttca caagtgggac ccccgcagcc
     3121 acctgcagcc ccttgctggg accccagagg ccacctacca gatgactgtg gagctcctgt
     3181 tcagggatat tctggaggag cccgaggagg atgagctcac agagaggctg agccagcacc
     3241 cgcacctgtg gctgtgcagg gcggccgcgg gttctcatca tcatcatcat catggtatgg
     3301 ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc
     3361 tcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac gcagtcagag
     3421 ctataggtcg gctgagctca gcgatcaaca ccgaaatgat gtacccagcg gatggtggtc
     3481 tgcgtggtta cactcacatg gcgctgaaag ttgatggcgg cggtcacctg tcctgttctt
     3541 tcgtgaccac ctaccgctcc aaaaagactg tcggcaacat taagatgcct gccattcatt
     3601 acgtcagcca ccgtctggag cgcctggagg agagcgataa cgaaatgttt gtcgtacagc
     3661 gtgaacacgc agttgccaag tttgtgggcc tgggtggtgg cggcggtacc ggagggagca
     3721 tgaactccct gatcaaggag aacatgcgta tgaaagtggt tctggaaggc tccgtaaacg
     3781 gccaccagtt caaatgcact ggtgaaggcg aaggcaaccc gtatatgggc acccagacta
     3841 tgcgtatcaa agtgatcgag ggtggtccgc tgccgtttgc gttcgacatc ctggcgacgt
     3901 cctttatgta tggctcccgt accttcatca aatatccgaa aggcatcccg gatttcttta
     3961 agcagtcctt cccggaaggt tttacctggg aacgtgtgac ccgttacgaa gacggcggcg
     4021 taattaccgt tatgcaagac acgtctctgg aggatggctg cctggtgtat cacgtgcagg
     4081 ttcgcggtgt taacttcccg agcaatggtg ctgtaatgca aaagaaaacc aaaggttggg
     4141 agcctacgga ctcccaactg actgaagagc agatcgcaga atttaaagag gctttctccc
     4201 tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg gtgatgcggt
     4261 ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
     4321 acggtgacgg cacaatcgac ttccctgagt tcctgattat gatggcaaga aaaatgaaat
     4381 acacagacag tgaagaagaa attagagaag cgttcggcgt gtttgataag gatggcaatg
     4441 gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag
     4501 atgaagaggt tgatgaaatg atcagggaag cagacagcga tggggatggt caggtaaact
     4561 acgaagagtt tgtacaaatg atgacagcga agtaatctag agggcccgtt taaacccgct
     4621 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4681 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4741 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4801 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     4861 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     4921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5101 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5221 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5281 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     5341 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5401 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5521 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5581 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5641 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5701 tttcggatct gatcagcacg tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     5761 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     5821 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     5881 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     5941 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     6001 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     6061 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     6121 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     6181 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     6241 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     6301 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     6361 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     6421 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     6481 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     6541 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     6601 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     6661 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     6721 cagcactcgt ccgagggcaa aggaatagca cgtgctacga gatttcgatt ccaccgccgc
     6781 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6841 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6901 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6961 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7021 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7081 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7141 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7201 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7981 tagcggttgg ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9181 tccccgaaaa gtgccacctg acgtc
//
