LOCUS       Exported                5332 bp ds-DNA     circular SYN 30-JAN-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 4, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5332
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    31..55
                     /label=TUPV2 Barcode
     promoter        82..431
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    90..108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    162..180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    198..216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    234..252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    270..288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    306..324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        476..494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      514..523
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
     CDS             520..1197
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     polyA_signal    1248..1472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1490..1514
                     /label=TUPV2 Barcode
     misc_feature    1538..1773
                     /label=cHS4
     misc_feature    1877..2112
                     /label=cHS4
     rep_origin      2241..2669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3580..4165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4336..5196)
                     /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"
     promoter        complement(5197..5301)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 caggcccttt catcttcact cgagtttact ccctatcagt gatagagaac gtatgaagag
      121 tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga
      181 acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat
      241 cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag
      301 tttactccct atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta
      361 taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt
      421 gtcttatact tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      481 cgactcacta tagggagtcc caagctggct agcgccacca tggatagcac tgagaacgtc
      541 atcaagccct tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      601 gagatcgagg gcgagggcga gggcaagccc tacgagggca cccagaccgc caagctccag
      661 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
      721 ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc
      781 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      841 acccaggact cctccctcca ggacggcacc ttcatctacc acgtgaagtt catcggcgtg
      901 aacttcccct ccgacggccc cgtaatgcag aagaaaactc tgggctggga gccctccacc
      961 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccacaaggc gctgaagctg
     1021 aagggcggcg gccactacct ggtggagttc aagtcaatct acatggccaa gaagcccgtg
     1081 aagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca caacgaggac
     1141 tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagtaggcg
     1201 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     1261 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1321 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1381 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagtcaatag
     1441 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcaggtcgaa gaactatttc
     1501 ctggctgtta cgcgagcgca tcgcgataag ctctagaggg acagcccccc cccaaagccc
     1561 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     1621 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     1681 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     1741 cagacacctg gggggatacg gggaaaaagc tttaggctga aagagagatt tagaatgaca
     1801 gaatcataga acggcctgcg tacggatggg gctctagctc tagagcttga tatcgaattc
     1861 ctgcagcccc gctagaggga cagccccccc ccaaagcccc cagggatgta attacgtccc
     1921 tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc
     1981 cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat
     2041 cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg
     2101 ggaaaaagct ttaggctgaa agagagattt agaatgacag aatcatagaa cggcctgggc
     2161 gcgcctggtg gcgaacgatc aacagacggg agaggactaa agtcttcgct gacagcgacc
     2221 ggactctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2641 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttaggctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct tcggaatcgt
     2941 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3001 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3121 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3181 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3241 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3301 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3361 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3421 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3481 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3541 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3601 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3661 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3721 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3781 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3841 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3901 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3961 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4021 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4081 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     4141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4441 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga
     4501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
