LOCUS       sequence-94230-e        8544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MESA target chain with CD4 ectodomain, no linker domain, G cleavage 
            sequence, and tTA-BFP fusion .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8544)
  AUTHORS   Daringer NM, Dudek RM, Schwarz KA, Leonard JN
  TITLE     Modular Extracellular Sensor Architecture for Engineering Mammalian 
            Cell-based Devices.
  JOURNAL   ACS Synth Biol. 2014 Mar 11.
  PUBMED    24611683
REFERENCE   2  (bases 1 to 8544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2014 Mar 11."
COMMENT     Sequence Label: CD4_LD 0_CS G_tTA-BFP
FEATURES             Location/Qualifiers
     source          1..8544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     709..729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2095..2115
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2116..2736,2737..3120)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2189..2208)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             join(3151..3153,3154..3156,3157..3870)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(3196..3217)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3991..4383
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      5291..5804
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5378..5397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5588..5609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6285..6304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6404..6426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6464..6482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6550..6654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6655..6723,6724..7515)
                     /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(6873..6892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7686..8274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8175..8194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(8514..8544,1..99)
                     /label=AAV2 ITR
ORIGIN
        1 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
       61 gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc
      121 catgctactt atctacgtag ccatgctcta gggatccggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      841 gccatcatgt gccgagccat ctctcttagg cgcttgctgc tgctgctgct gcagctgtca
      901 caactcctag ctgtcactca agggaagacg ctggtgctgg ggaaggaagg ggaatcagca
      961 gaactgccct gcgagagttc ccagaagaag atcacagtct tcacctggaa gttctctgac
     1021 cagaggaaga ttctggggca gcatggcaaa ggtgtattaa ttagaggagg ttcgccttcg
     1081 cagtttgatc gttttgattc caaaaaaggg gcatgggaga aaggatcgtt tcctctcatc
     1141 atcaataaac ttaagatgga agactctcag acttatatct gtgagctgga gaacaggaaa
     1201 gaggaggtgg agttgtgggt gttcaaagtg accttcagtc cgggtaccag cctgttgcaa
     1261 gggcagagcc tgaccctgac cttggatagc aactctaagg tctctaaccc cttgacagag
     1321 tgcaaacaca aaaagggtaa agttgtcagt ggttccaaag ttctctccat gtccaaccta
     1381 agggttcagg acagcgactt ctggaactgc accgtgaccc tggaccagaa aaagaactgg
     1441 ttcggcatga cactctcagt gctgggtttt cagagcacag ctatcacggc ctataagagt
     1501 gagggagagt cagcggagtt ctccttccca ctcaactttg cagaggaaaa cgggtgggga
     1561 gagctgatgt ggaaggcaga gaaggattct ttcttccagc cctggatctc cttctccata
     1621 aagaacaaag aggtgtccgt acaaaagtcc accaaagacc tcaagctcca gctgaaggaa
     1681 acgctcccac tcaccctcaa gataccccag gtctcgcttc agtttgctgg ttctggcaac
     1741 ctgactctga ctctggacaa agggacactg catcaggaag tgaacctggt ggtgatgaaa
     1801 gtggctcagc tcaacaatac tttgacctgt gaggtgatgg gacctacctc tcccaagatg
     1861 agactgaccc tgaagcagga gaaccaggag gccagggtct ctgaggagca gaaagtagtt
     1921 caagtggtgg cccctgagac agggctgtgg cagtgtctac tgagtgaagg tgataaggtc
     1981 aagatggact ccaggatcca ggttttatcc agagggaagc ttttttgggc actggtcgtg
     2041 gttgctggag tcctgttttg ttatggcttg ctagtgacag tggctctttg tgttgaaaac
     2101 ctgtattttc agggtatgtc tagattagat aaaagtaaag tgattaacag cgcattagag
     2161 ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt
     2221 gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta
     2281 gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg
     2341 caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat
     2401 ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat
     2461 caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc
     2521 gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa
     2581 gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa
     2641 ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc
     2701 ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa
     2761 aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga cgcccccgaa
     2821 gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac gcgcagactg
     2881 tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
     2941 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
     3001 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
     3061 gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtgga
     3121 ggtaccggcg gaggctccgg tggtggctct atggtgagca agggcgagga gctgttcacc
     3181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3241 aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc
     3301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag
     3361 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta caagacccgc
     3481 gccgaggtga agttcgaggg cgacacccta gtgaaccgca tcgagctgaa gggcgtcgac
     3541 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac
     3601 atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3661 aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3721 gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt gctgagcaaa
     3781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc cgccgggatc
     3841 actctcggca tggacgagct gtacaagtaa tagcggccgc caccgcggtg gagctgttta
     3901 aacctcgagc aattcacccc accagtgcag gctgcctatc agaaagtggt ggctggtgtg
     3961 gctaatgccc tggcccacaa gtatcactaa gctcgctttc ttgctgtcca atttctatta
     4021 aaggttcctt tgttccctaa gtccaactac taaactgggg atattatgaa gggccttgag
     4081 catctggatt ctgcctaata aaaaacattt attttcattg caatgatgta tttaaattat
     4141 ttctgaatat tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa
     4201 tgaagagcta gttcaaacct tgggaaaata cactatatct taaactccat gaaagaaggt
     4261 gaggctgcaa acagtaatgc acattggcaa cagcccctga tgcctatgcc ttattcatcc
     4321 ctcagaaaag gattcaagta gaggcttgat ttggaggtta aagttttgct atgctgtatt
     4381 ttacattact tattgtttta gctgtcctca tgaatgtctt ttcactaccc atttgcttat
     4441 cctgcatctc tcagccttga ctccactcag ttctcttgct tagagatacc acctttcccc
     4501 tgaagtgttc cttccatgtt ttacggcgag atggtttctc ctcgcctggc cactcagcct
     4561 tagttgtctc tgttgtctta tagaggtcta cttgaagaag gaaaaacagg gggcatggtt
     4621 tgactgtcct gtgagccctt cttccctgcc tcccccactc acagtgaccc ggaatccctc
     4681 gacatggcag tctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     4741 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4801 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4861 agcgagcgcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4921 ttgcgcagcc tgaatggcga atggaattcc agacgattga gcgtcaaaat gtaggtattt
     4981 ccatgagcgt ttttcctgtt gcaatggctg gcggtaatat tgttctggat attaccagca
     5041 aggccgatag tttgagttct tctactcagg caagtgatgt tattactaat caaagaagta
     5101 ttgcgacaac ggttaatttg cgtgatggac agactctttt actcggtggc ctcactgatt
     5161 ataaaaacac ttctcaggat tctggcgtac cgttcctgtc taaaatccct ttaatcggcc
     5221 tcctgtttag ctcccgctct gattctaacg aggaaagcac gttatacgtg ctcgtcaaag
     5281 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5341 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5401 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5461 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5521 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5581 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5641 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5701 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaat atttgcttat
     5761 acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtacatat gattgacatg
     5821 ctagttttac gattaccgtt catcgattct cttgtttgct ccagactctc aggcaatgac
     5881 ctgatagcct ttgtagagac ctctcaaaaa tagctaccct ctccggcatg aatttatcag
     5941 ctagaacggt tgaatatcat attgatggtg atttgactgt ctccggcctt tctcacccgt
     6001 ttgaatcttt acctacacat tactcaggca ttgcatttaa aatatatgag ggttctaaaa
     6061 atttttatcc ttgcgttgaa ataaaggctt ctcccgcaaa agtattacag ggtcataatg
     6121 tttttggtac aaccgattta gctttatgct ctgaggcttt attgcttaat tttgctaatt
     6181 ctttgccttg cctgtatgat ttattggatg ttggaattcc tgatgcggta ttttctcctt
     6241 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     6301 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6361 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     6421 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     6481 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7801 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     7861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8341 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8401 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8461 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag cagctgcgcg
     8521 ctcgctcgct cactgaggcc gccc
//
