LOCUS       Exported                6086 bp ds-DNA     circular SYN 20-MAY-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  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..6086
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    83..245
                     /label=Entire ZF43x6-Compact Promoter System
                     /note="From:Nat Commun. 2020 Feb 7;11(1):779. doi: 
                     10.1038/s41467-019-14147-5. (PMID 32034124)"
                     /note="color: #333399"
     misc_feature    101..109
                     /label=ZF43 Binding Site 1
                     /note="Flanked by g/t"
                     /note="color: #a6acb3"
     misc_feature    117..124
                     /label=ZF43 Binding Site 2
                     /note="Flanked by a/c"
                     /note="color: #a6acb3"
     misc_feature    131..139
                     /label=ZF43 Binding Site 3
                     /note="Flanked by g/a"
                     /note="color: #a6acb3"
     misc_feature    146..154
                     /label=ZF43 Binding Site 4
                     /note="Flanked by a/c"
                     /note="color: #a6acb3"
     misc_feature    161..169
                     /label=ZF43 Binding Site 5
                     /note="Flanked by g/t"
                     /note="color: #a6acb3"
     misc_feature    176..184
                     /label=ZF43 Binding Site 6
                     /note="Flanked by a/c"
                     /note="color: #a6acb3"
     TATA_signal     192..216
                     /label=YB_TATA Minimal Promoter
                     /note="color: #ffffff"
     intron          357..929
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
                     /note="color: #------"
     CDS             1015..2548
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /note="This feature has 2 segments:
                        1: 1015 .. 1062 / #993366 / signal peptide
                        2: 1063 .. 2548 / #993366"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     3'UTR           2551..2650
                     /label=VSV-G 3'UTR (must keep with VSV-G CDS if cloning)
                     /note="color: #008000"
     polyA_signal    2842..2897
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
                     /note="color: #a6acb3"
     primer_bind     complement(3255..3271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    3279..3295
                     /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(3303..3333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 3303 .. 3309 / #ffffff / -10
                        2: 3310 .. 3327 / #ffffff
                        3: 3328 .. 3333 / #ffffff / -35"
     protein_bind    3348..3369
                     /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(3657..4245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4416..5276)
                     /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: 4416 .. 5207 / #ccffcc
                        2: 5208 .. 5276 / #ccffcc / signal sequence
                     Cleavage site after base 5207"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5277..5381)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(5407..5862)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     6004..6020
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        6030..6048
                     /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 ttagatctga attacgatcg gagtgaggat aattagagtg
      121 aggacgatcg gagtgaggaa aattagagtg aggacgatcg gagtgaggat aattagagtg
      181 aggacgatcc ctctagaggg tatataatgg gggccactag tctactacca gagctcatcg
      241 ctagccgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      301 atagaagaca ccgggaccga tccagcctcc ggtcgaccga tcctgagaac ttcagggtga
      361 gtttggggac ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag
      421 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac
      481 cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta
      541 ttttcttttc attttctgta actttttcgt taaactttag cttgcatttg taacgaattt
      601 ttaaattcac ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc
      661 aaggcaatca gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa
      721 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
      781 ttggtagaaa caactacacc ctggtcatca tcctgccttt ctctttatgg ttacaatgat
      841 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
      901 atgttcatgc cttcttctct ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt
      961 ctcatcattt tggcaaagaa ttcctcgacg gatccctcga ggaattctga cactatgaag
     1021 tgccttttgt acttagcctt tttattcatt ggggtgaatt gcaagttcac catagttttt
     1081 ccacacaacc aaaaaggaaa ctggaaaaat gttccttcta attaccatta ttgcccgtca
     1141 agctcagatt taaattggca taatgactta ataggcacag ccttacaagt caaaatgccc
     1201 aagagtcaca aggctattca agcagacggt tggatgtgtc atgcttccaa atgggtcact
     1261 acttgtgatt tccgctggta tggaccgaag tatataacac attccatccg atccttcact
     1321 ccatctgtag aacaatgcaa ggaaagcatt gaacaaacga aacaaggaac ttggctgaat
     1381 ccaggcttcc ctcctcaaag ttgtggatat gcaactgtga cggatgccga agcagtgatt
     1441 gtccaggtga ctcctcacca tgtgctggtt gatgaataca caggagaatg ggttgattca
     1501 cagttcatca acggaaaatg cagcaattac atatgcccca ctgtccataa ctctacaacc
     1561 tggcattctg actataaggt caaagggcta tgtgattcta acctcatttc catggacatc
     1621 accttcttct cagaggacgg agagctatca tccctgggaa aggagggcac agggttcaga
     1681 agtaactact ttgcttatga aactggaggc aaggcctgca aaatgcaata ctgcaagcat
     1741 tggggagtca gactcccatc aggtgtctgg ttcgagatgg ctgataagga tctctttgct
     1801 gcagccagat tccctgaatg cccagaaggg tcaagtatct ctgctccatc tcagacctca
     1861 gtggatgtaa gtctaattca ggacgttgag aggatcttgg attattccct ctgccaagaa
     1921 acctggagca aaatcagagc gggtcttcca atctctccag tggatctcag ctatcttgct
     1981 cctaaaaacc caggaaccgg tcctgctttc accataatca atggtaccct aaaatacttt
     2041 gagaccagat acatcagagt cgatattgct gctccaatcc tctcaagaat ggtcggaatg
     2101 atcagtggaa ctaccacaga aagggaactg tgggatgact gggcaccata tgaagacgtg
     2161 gaaattggac ccaatggagt tctgaggacc agttcaggat ataagtttcc tttatacatg
     2221 attggacatg gtatgttgga ctccgatctt catcttagct caaaggctca ggtgttcgaa
     2281 catcctcaca ttcaagacgc tgcttcgcaa cttcctgatg atgagagttt attttttggt
     2341 gatactgggc tatccaaaaa tccaatcgag cttgtagaag gttggttcag tagttggaaa
     2401 agctctattg cctctttttt ctttatcata gggttaatca ttggactatt cttggttctc
     2461 cgagttggta tccatctttg cattaaatta aagcacacca agaaaagaca gatttataca
     2521 gacatagaga tgaaccgact tggaaagtaa ctcaaatcct gcacaacaga ttcttcatgt
     2581 ttggaccaaa tcaacttgtg ataccatgct caaagaggcc tcaattatat ttgagttttt
     2641 aatttttatg aaaaaaaaaa aaaaaaacgg aattcctcga gggatccgtc gaggaattca
     2701 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     2761 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     2821 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2881 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     2941 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     3001 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     3061 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3121 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3181 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ggatcggccg
     3241 caattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3301 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3361 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3421 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3481 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3541 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3601 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3661 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3721 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3781 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3841 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3901 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3961 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4021 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4081 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4141 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4201 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4261 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4321 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4381 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4441 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4501 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4561 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4621 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4681 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4741 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4801 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4861 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4921 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4981 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5041 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5101 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5161 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5221 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5281 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5341 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5401 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5461 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5521 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5581 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5641 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5701 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5761 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5821 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     5881 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     5941 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6001 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggagc
     6061 tccaccgcgg tggcggccgc tctaga
//
