LOCUS       pcDNA5TO_HsTPC2-        8581 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8581)
  AUTHORS   Medical Sciences Division
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8581
                     /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: #993366; direction: RIGHT"
     CDS             3270..3977
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     polyA_signal    4009..4233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4279..4707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4721..5050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4901..5036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             5099..6124
                     /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    6254..6375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(6424..6440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    6448..6464
                     /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(6472..6478,6479..6496,6497..6502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 6472 .. 6478 / #ffffff / -10
                        2: 6479 .. 6496 / #ffffff
                        3: 6497 .. 6502 / #ffffff / -35"
     protein_bind    6517..6538
                     /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(6826..7414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(join(7585..8376,8377..8445))
                     /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: 7585 .. 8376 / #ccffcc
                        2: 8377 .. 8445 / #ccffcc / signal sequence
                     Cleavage site after base 8376"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8446..8550)
                     /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 tgagcaaggg cgaggaggat aacatggcca
     3301 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     3361 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     3421 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     3481 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     3541 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     3601 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     3661 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     3721 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     3781 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     3841 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     3901 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     3961 acgagctgta caagtagggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     4021 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4081 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4141 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4201 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4261 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4321 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4381 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4441 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4501 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4561 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4621 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4681 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4741 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4801 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4861 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4921 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4981 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5041 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat
     5101 gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     5161 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     5221 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     5281 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     5341 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     5401 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     5461 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     5521 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     5581 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5641 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5701 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5761 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5821 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     5881 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5941 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     6001 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     6061 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     6121 atagcacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6181 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6241 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6301 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6361 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6421 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6481 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6541 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6601 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6661 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6721 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6781 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6841 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6901 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6961 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7021 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7081 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7141 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7201 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7261 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7321 tggtagctct tgatccggca aacaaaccac cgctggtagc ggttggtttt ttgtttgcaa
     7381 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7441 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7501 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8101 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8461 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8521 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8581 c
//
