LOCUS       TMEM_PhoCl2c_rtt        9806 bp DNA     circular SYN 21-FEB-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 10, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..478
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    561..1613
                     /label=TMEM115
     misc_feature    1641..2366
                     /label=PhoCl2c
     CDS             2397..3101
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     misc_feature    3117..3815
                     /label=mscarlet
     polyA_signal    3887..4008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      4123..4169
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4199..4335
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4380..4451
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4677..5265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5765..6974
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             6978..8006
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    8033..8081
                     /note="synthetic polyadenylation signal"
     CDS             join(8347..8415,8416..9207)
                     /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"
     terminator      9220..9356
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9508..9579
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      9584..9630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9648..9696
                     /note="synthetic polyadenylation signal"
     enhancer        join(9777..9806,1..274)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
       61 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      121 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      181 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      241 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
      301 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      361 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      421 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      481 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
      541 gcggccgcgc tagcgccacc atgcaacgtg ccctgccagg cgcccgccag cacttggggg
      601 ccattctggc cagcgccagc gtggtggtga aggctctgtg tgcggcggta ctattcctct
      661 acctgctctc cttcgccgtg gacacaggct gcctggcggt caccccgggc tacctctttc
      721 ctcccaactt ctggatctgg accctggcca cccatgggct gatggagcag catgtgtggg
      781 acgtggccat cagcctgaca acggtggtgg tggccgggcg tttgctggag cccctctggg
      841 gggccttgga gctgctcatc ttcttctcag tggtgaatgt gtctgtaggg ctgctggggg
      901 ccttcgccta cctcctcacc tacatggctt ccttcaacct ggtctacctg ttcactgtcc
      961 gtatccacgg cgccttgggc ttcctaggtg gcgtcctggt ggcactcaag caaaccatgg
     1021 gggactgtgt ggtcctgcga gtgccccagg tgcgcgtcag tgtgatgccc atgctgctgc
     1081 tggcgctgct gctcctgctg cggctcgcca cgctgctcca gagcccggcg ctggcttcct
     1141 atggcttcgg gctgctctcc agttgggtat atcttcgctt ctaccagcgc catagccggg
     1201 gccgagggga catggctgac cactttgctt tcgccacttt cttccctgag atcctgcagc
     1261 ctgtggtggg tttgctggcg aacttggtgc acagcctcct ggtgaaggta aagatatgcc
     1321 agaagacggt gaagcgctac gatgtgggtg ccccatcctc catcaccatc agcctgccag
     1381 gcacagaccc tcaagacgcc gagcggagaa ggcaactggc cctgaaggca ctcaatgagc
     1441 ggctgaagag agtggaagac cagtccatct ggcccagcat ggatgatgat gaagaggagt
     1501 ctggggccaa ggtggacagc cccctgccct cagacaaagc tcccacaccc ccagggaagg
     1561 gggctgcccc agaatccagt ctaatcacct tcgaggcagc tcccccgacg ctgtgtacaa
     1621 gtccggactc agatctcgag gtcatacccg actacttcaa gcagagcttc cccgagggct
     1681 acagctggga gagatccatg acatacgagg acggcggcat ctgtatcgcc accaacgaca
     1741 tcaccatgga aggcgacagc ttcatcaaca agatccactt tcagggcacg aacttccctc
     1801 caaacggccc cgtgatgcag aagagaacag ttggctggga agccagcacc gagaagatgt
     1861 acgagcgaga tggcgtgctg aagggcgacg tgaaaatgaa gctgctgctg aaaggcggcg
     1921 gacactacag aggcgactac cggaccacct acaaagtgaa gcagaaaccc gtgaagctgc
     1981 ccgactgcca cttcgtggat caccggatcg agatcctgag ccacgacaag gactacaaca
     2041 aagtcaagct gtacgagcac gccgtggcca agacctccac cgattctatg gacgagctgt
     2101 acaaaggcgg cagcggcggc atggtgtcta agggcgaaga gacaatcacc agcgtgatca
     2161 agcccgacat gaagaacaag ctgcggatgg aaggcaacgt gaacggccac gcctttgtga
     2221 tcgaaggcga aggcagcggc aagccctttg agggcatcca gaccatcgac ctggaagtga
     2281 aagaaggcgc ccctctgcct ttcgcctacg acattctgac caccgccttc cactacggca
     2341 accgggtgtt cacaaagtac cctagaggcg gaggcggcac aaagcttggt agtggcatgt
     2401 ctagactgga caagagcaaa gtcataaacg gagctctgga attactcaat ggtgtcggta
     2461 tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt
     2521 actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca
     2581 ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca
     2641 acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg
     2701 gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc
     2761 agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac
     2821 tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta
     2881 ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag
     2941 ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa
     3001 agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag
     3061 ccgatgccct tgacgatttt gaccttgaca tgctccccgg ggaattcggt agtggcatgg
     3121 tttctaaagg cgaggccgtg atcaaagagt tcatgcggtt caaggtgcac atggaaggca
     3181 gcatgaacgg ccacgagttc gagatcgaag gcgaaggcga gggcagacct tatgagggaa
     3241 cacagaccgc caagctgaaa gtgaccaaag gcggccctct gcctttcagc tgggacattc
     3301 tgagccctca gtttatgtac ggcagccggg ccttcacaaa gcaccctgcc gatattcccg
     3361 actactacaa gcagagcttc cccgagggct tcaagtggga gagagtgatg aacttcgagg
     3421 acggcggagc cgtgaccgtg acacaggata caagcctgga agatggcacc ctgatctaca
     3481 aagtgaagct gcggggcacc aactttccac ctgatggccc cgtgatgcag aaaaagacca
     3541 tgggctggga agccagcacc gagagactgt atcctgagga tggcgtgctg aagggcgaca
     3601 tcaagatggc cctgagactg aaggatggcg gcagatacct ggccgacttc aagaccacct
     3661 acaaggccaa gaaacccgtg cagatgcctg gcgcctacaa cgtggacaga aagctggaca
     3721 tcaccagcca caacgaggac tacaccgtgg tggaacagta cgagcggagc gaaggcagac
     3781 actctacagg cggaatggac gagctgtaca agtaaccgcg gggatccgca ggcctctgct
     3841 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     3901 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3961 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     4021 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     4081 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     4141 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     4201 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     4261 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     4321 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     4381 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     4441 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     4501 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     4561 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     4621 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4681 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4741 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4801 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4861 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4921 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4981 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5041 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5101 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5161 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5221 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5281 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5341 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5401 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5461 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     5521 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     5581 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     5641 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     5701 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     5761 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     5821 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     5881 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     5941 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     6001 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     6061 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     6121 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     6181 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     6241 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     6301 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     6361 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     6421 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     6481 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     6541 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     6601 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg
     6661 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     6721 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     6781 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     6841 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     6901 caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg
     6961 cttttttgtt agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     7021 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     7081 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     7141 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga
     7201 ttccggaagt gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc
     7261 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     7321 cggtcgcgga ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg
     7381 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     7441 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     7501 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     7561 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     7621 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     7681 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     7741 cggagcttgc aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac
     7801 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     7861 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     7921 ccgccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     7981 gcactcgtcc gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat
     8041 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     8101 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     8161 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     8221 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     8281 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     8341 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     8401 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     8461 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     8521 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     8581 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     8641 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     8701 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     8761 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     8821 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     8881 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     8941 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     9001 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     9061 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     9121 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     9181 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     9241 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     9301 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     9361 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     9421 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     9481 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     9541 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     9601 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     9661 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     9721 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     9781 acataactta cggtaaatgg cccgcc
//
