LOCUS       sequence-150732-        7929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7929)
  AUTHORS   Cetin A, Callaway EM
  TITLE     Optical control of retrogradely infected neurons using 
            drug-regulated ""TLoop"" lentiviral vectors.
  JOURNAL   J Neurophysiol. 2014 May;111(10):2150-9. doi: 10.1152/jn.00495.2013.
            Epub 2014 Feb 26.
  PUBMED    24572099
REFERENCE   2  (bases 1 to 7929)
  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"; doi: 
            "10.1152/jn.00495.2013"; journalName: "J Neurophysiol"; date: 
            "2014-05"; volume: "111"; issue: "10"; pages: "2150-9"
COMMENT     Sequence Label: pEIAV-TLoop-DsRedx
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(1913..1931,1932..1954,1955..1973,1974..1996,1997..2015,
                     2016..2038,2039..2057,2058..2080,2081..2099,2100..2122,
                     2123..2141,2142..2164,2165..2183)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2216..2254
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          2409..2638
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2621..2640
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2693..2702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2699..3373
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2829..2847)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2879..2899)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3097..3116
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3291..3314
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3383..3436
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3455..4072
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     misc_feature    4366..4954
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4419..4439)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4837..4848)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      5351..5806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5438..5457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5648..5669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5875..5894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5994..6016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6054..6072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6140..6244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6245..6313,6314..7105)
                     /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(6463..6482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7276..7864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7765..7784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
       61 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
      121 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgggcactc agattctgcg gtctgagtcc cttctctgct
      781 gggctgaaaa ggcctttgta ataaatataa ttctctactc agtccctgtc tctagtttgt
      841 ctgttcgaga tcctacagtt ggcgcccgaa cagggacctg agaggggcgc agaccctacc
      901 tgttgaacct ggctgatcgt aggatccccg ggacagcaga ggagaactta cagaagtctt
      961 ctggaggtgt tcctggccag aacacaggag gacaggtaag tagggagacc ctttgacatg
     1021 gagcaaggcg ctcaagaagt tagagaaggt gacggtacaa gggtctcaga aattaactac
     1081 tggtaactgt aattgggcgc taagtctagt agacttattt catgatacca actttgtaaa
     1141 agaaaaggac tggcagctga gggatgtcat tccattgctg gaagatgtaa ctcagacgct
     1201 ggaattcggg tattgtagaa agggcaaata ggaccttgaa agagaagatt caaagtcata
     1261 gagacaacac tcaaacactg gaggcagctt tacaacttgc tctcattact tgtaacaaag
     1321 ggagggaaag tatgggagga cagacaccat gggaagtatt tatcactaat caagcacaag
     1381 taatacatga gaaactttta ctacagcaag cacaatcctc caaaaaattt tgtttttaca
     1441 aaatccctgg tgaacatgat tggaagggac ctactagggt gctgtggaag ggtgatggtg
     1501 cagtagtagt taatgatgaa ggaaagggaa taattgctgt accattaacc aggactaagt
     1561 tactaataaa gccaaattga gtattgttgc atgcatttga tatatgggat tatttggtaa
     1621 aggggtaaca tggtcagcat cgcattctat ggggggatcc cagggggaat ctcaacccct
     1681 attacccaac agtcagaaaa atctaagtgt gaggagaaca caatgtttca accttattgt
     1741 tataataatg acagtaagaa cagcatggca gaatcgaagg aagcaagaga ccaagaaatg
     1801 aacctgaaag aagaatctaa agaagaaaaa agaagaaatg actggtggaa aaaagctgca
     1861 gagggccctt aatgcagtgg cctttgcggc cgccaactct cgagtttacc actccctatc
     1921 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
     1981 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     2041 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     2101 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     2161 ccactcccta tcagtgatag agaaaagtga aagtcgagct cggtacccgg gtcgaggtag
     2221 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2281 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     2341 cggccccgaa ttcaccacgt ggccgcatcg attctagaat tcgctgtctg cgagggccag
     2401 ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa gattgtcagt
     2461 ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt tgagggtggc
     2521 cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt gtggcaggct
     2581 tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct ctccacaggt
     2641 gtccactccc aggtccaact gcagcccaag cggatcctct agagtcgacg ccgccaccat
     2701 ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca tggagggctc
     2761 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2821 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct
     2881 gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg acatccccga
     2941 ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3001 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggctcct tcatctacaa
     3061 ggtgaagttc atcggcgtga acttcccctc cgacggcccc gtaatgcaga agaagactat
     3121 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat
     3181 ccacaaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca agtccatcta
     3241 catggccaag aagcccgtgc agctgcccgg ctactactac gtggactcca agctggacat
     3301 cacctcccac aacgaggact acaccatcgt ggagcagtac gagcgcgccg agggccgcca
     3361 ccacctgttc ctgctagccg cggagggccg cggcagcctg ctgacctgcg gcgacgtgga
     3421 ggagaacccc ggccccgccc ccggcagcgg tacctccaga ctggacaaga gcaaagtcat
     3481 caactctgcc ttggagctcc tgaatgaagt tggcattgag ggcttgacca ccaggaagct
     3541 ggcccagaag ctgggtgtgg agcagcctac cctgtactgg catgtgaaga acaagagggc
     3601 tctgcttgat gccctggcca ttgagatgtt ggacaggcac cacacccact tctgccctct
     3661 ggaaggggag tcctggcagg acttcctgag gaacaatgcc aagagcttca gatgtgcctt
     3721 gctctcccac cgggatggtg ccaaagttca cttgggcacc aggcctacag agaagcagta
     3781 tgagaccctg gagaaccagc tggcattcct gtgccaacaa ggcttctccc tggaaaatgc
     3841 cttgtatgcc ctctctgctg tgggccactt caccttgggc tgtgtgctgg aggaccagga
     3901 gcaccaagtt gccaaggagg agagggagac ccccaccact gactccatgc caccactgct
     3961 gcggcaagct attgagttgt ttgaccacca aggggctgag cctgcattcc tttttggcct
     4021 ggaactgatc atctgtggcc tggaaaagca gctgaaatgt gagtctggct ctcacgtgcc
     4081 caaaaagaga aagcacgtgc ctgctgatgc cttggatgat tttgacctgg acatgctgcc
     4141 tgctgatgcc ctggatgact ttgatttgga catgctccct gctgatgcac ttgatgattt
     4201 tgacctggat atgctgtgat ccggagtcta catttgaatg tcaataaaac aaaatggtgg
     4261 ggcaatcatc tacatttcat gggatatgtg attactagtt caggtgtatt gccacaagac
     4321 aaacatgtta agaaaatttc ccgttatttg cactctgttc ctgttaatca acctctggat
     4381 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4441 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4501 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4561 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4621 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4681 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4741 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     4801 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4861 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4921 acgagtcgga tctccctttg ggccgcctcc ccgcctgttt cgcctcggcg tccggtccgt
     4981 gttgcttggt cttcacctgt gcagacttgc gaaccatgga ttccaccgtg aactttgtct
     5041 cctggcatgc aaatcgtcaa cttggcatgc caaatcgatt agaaaaacaa ggggggaact
     5101 gtggggtttt tatgaggggt tttatacaat tgggcactca gattctgcgg tctgagtccc
     5161 ttctctgctg ggctgaaaag gcctttgtaa taaatataat tctctactca gtccctgtct
     5221 ctagtttgtc tgttcgagat cctacagagc tcatgccttg gttaattaag gagatccggg
     5281 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5341 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5401 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5461 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5521 ttccgattta gagctttacg gcacctcgac cgcaaaaaac ttgatttggg tgatggttca
     5581 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5641 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5701 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5761 caaatattta acgcgaattt taacaaaata ttaacgttta caatttcgcc tgatgcggta
     5821 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     5881 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5941 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6001 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     6061 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6181 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6241 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6301 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6361 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6421 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6481 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6541 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6601 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6661 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6721 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6781 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6841 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6901 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6961 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7021 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7081 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7141 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7201 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7261 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7321 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7381 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     7441 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7501 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7561 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7621 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7681 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7741 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7801 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7861 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7921 catggctcg
//
