LOCUS       Exported                6302 bp ds-DNA     circular SYN 19-MAY-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  AUTHORS   Dr John and Anne Chong Lab for Functional Genomics
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 21, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6302
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        83..461
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
                     /note="color: #ffffff; direction: RIGHT"
                     /note="color: #00ff00"
     protein_bind    91..109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    127..145
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(1)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    163..181
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(2)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    199..217
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(3)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    235..253
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(4)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    271..289
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(5)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     protein_bind    307..325
                     /gene="tetO"
                     /label=tet operator
                     /label=tet operator(6)
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
                     /note="color: #ffc0cb"
     primer_bind     370..394
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
                     /note="color: black; sequence: AGCTCGTTTAGTGAACCGTCAGATC"
                     /note="color: #ffc0cb; direction: RIGHT"
     intron          573..1145
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
                     /note="color: #------"
     CDS             1231..2764
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /note="This feature has 2 segments:
                        1: 1231 .. 1278 / #993366 / signal peptide
                        2: 1279 .. 2764 / #993366"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     3'UTR           2767..2866
                     /label=VSV-G 3'UTR (must keep with VSV-G CDS if cloning)
                     /note="color: #008000"
     polyA_signal    3058..3113
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(3471..3487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    3495..3511
                     /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(3519..3549)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 3519 .. 3525 / #ffffff / -10
                        2: 3526 .. 3543 / #ffffff
                        3: 3544 .. 3549 / #ffffff / -35"
     protein_bind    3564..3585
                     /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(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4632..5492)
                     /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: 4632 .. 5423 / #ccffcc
                        2: 5424 .. 5492 / #ccffcc / signal sequence
                     Cleavage site after base 5423"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5493..5597)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(5623..6078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     6220..6236
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        6246..6264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 gagcttggcc cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
       61 tgtccaacat taccgccatg ttgagtttac tccctatcag tgatagagaa cgtatgaaga
      121 gtttactccc tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag
      181 aacgtataag gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta
      241 tcagtgatag agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca
      301 gtttactccc tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta
      361 taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt
      421 gtcttatacc aactttccgt accacttcct accctcgtaa acgtttagtg aaccgtcaga
      481 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      541 gcctccggtc gaccgatcct gagaacttca gggtgagttt ggggaccctt gattgttctt
      601 tctttttcgc tattgtaaaa ttcatgttat atggaggggg caaagttttc agggtgttgt
      661 ttagaatggg aagatgtccc ttgtatcacc atggaccctc atgataattt tgtttctttc
      721 actttctact ctgttgacaa ccattgtctc ctcttatttt cttttcattt tctgtaactt
      781 tttcgttaaa ctttagcttg catttgtaac gaatttttaa attcactttt gtttatttgt
      841 cagattgtaa gtactttctc taatcacttt tttttcaagg caatcagggt atattatatt
      901 gtacttcagc acagttttag agaacaattg ttataattaa atgataaggt agaatatttc
      961 tgcatataaa ttctggctgg cgtggaaata ttcttattgg tagaaacaac tacaccctgg
     1021 tcatcatcct gcctttctct ttatggttac aatgatatac actgtttgag atgaggataa
     1081 aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc ttctctttcc
     1141 tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaagaattcc
     1201 tcgacggatc cctcgaggaa ttctgacact atgaagtgcc ttttgtactt agccttttta
     1261 ttcattgggg tgaattgcaa gttcaccata gtttttccac acaaccaaaa aggaaactgg
     1321 aaaaatgttc cttctaatta ccattattgc ccgtcaagct cagatttaaa ttggcataat
     1381 gacttaatag gcacagcctt acaagtcaaa atgcccaaga gtcacaaggc tattcaagca
     1441 gacggttgga tgtgtcatgc ttccaaatgg gtcactactt gtgatttccg ctggtatgga
     1501 ccgaagtata taacacattc catccgatcc ttcactccat ctgtagaaca atgcaaggaa
     1561 agcattgaac aaacgaaaca aggaacttgg ctgaatccag gcttccctcc tcaaagttgt
     1621 ggatatgcaa ctgtgacgga tgccgaagca gtgattgtcc aggtgactcc tcaccatgtg
     1681 ctggttgatg aatacacagg agaatgggtt gattcacagt tcatcaacgg aaaatgcagc
     1741 aattacatat gccccactgt ccataactct acaacctggc attctgacta taaggtcaaa
     1801 gggctatgtg attctaacct catttccatg gacatcacct tcttctcaga ggacggagag
     1861 ctatcatccc tgggaaagga gggcacaggg ttcagaagta actactttgc ttatgaaact
     1921 ggaggcaagg cctgcaaaat gcaatactgc aagcattggg gagtcagact cccatcaggt
     1981 gtctggttcg agatggctga taaggatctc tttgctgcag ccagattccc tgaatgccca
     2041 gaagggtcaa gtatctctgc tccatctcag acctcagtgg atgtaagtct aattcaggac
     2101 gttgagagga tcttggatta ttccctctgc caagaaacct ggagcaaaat cagagcgggt
     2161 cttccaatct ctccagtgga tctcagctat cttgctccta aaaacccagg aaccggtcct
     2221 gctttcacca taatcaatgg taccctaaaa tactttgaga ccagatacat cagagtcgat
     2281 attgctgctc caatcctctc aagaatggtc ggaatgatca gtggaactac cacagaaagg
     2341 gaactgtggg atgactgggc accatatgaa gacgtggaaa ttggacccaa tggagttctg
     2401 aggaccagtt caggatataa gtttccttta tacatgattg gacatggtat gttggactcc
     2461 gatcttcatc ttagctcaaa ggctcaggtg ttcgaacatc ctcacattca agacgctgct
     2521 tcgcaacttc ctgatgatga gagtttattt tttggtgata ctgggctatc caaaaatcca
     2581 atcgagcttg tagaaggttg gttcagtagt tggaaaagct ctattgcctc ttttttcttt
     2641 atcatagggt taatcattgg actattcttg gttctccgag ttggtatcca tctttgcatt
     2701 aaattaaagc acaccaagaa aagacagatt tatacagaca tagagatgaa ccgacttgga
     2761 aagtaactca aatcctgcac aacagattct tcatgtttgg accaaatcaa cttgtgatac
     2821 catgctcaaa gaggcctcaa ttatatttga gtttttaatt tttatgaaaa aaaaaaaaaa
     2881 aaacggaatt cctcgaggga tccgtcgagg aattcactcc tcaggtgcag gctgcctatc
     2941 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     3001 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     3061 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3121 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3181 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3241 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3301 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3361 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3421 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     3481 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3541 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3601 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     5641 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5701 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5761 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5821 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5881 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5941 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     6001 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     6061 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6121 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6181 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     6241 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     6301 ga
//
