LOCUS       Exported                5249 bp DNA     circular SYN 30-JUN-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5249)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5249)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5249)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5249)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5249
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    58..187
                     /label=L-ITR
                     /note="L-ITR"
     polyA_signal    243..291
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     misc_feature    305..396
                     /label=pause site
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal
                     fromthe human alpha-2 globin gene"
     promoter        432..879
                     /label=hSyn promoter
                     /note="hSyn promoter"
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    674..681
                     /label=cre
                     /note="cre"
     regulatory      888..893
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="Kozak sequence"
                     /note="vertebrate consensus sequence for strong
                     initiationof translation (Kozak, 1987)"
     CDS             894..1640
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    1653..2241
                     /label=WPRE
                     /note="WPRE"
                     /note="woodchuck hepatitis virus
                     posttranscriptionalregulatory element"
     CDS             complement(2124..2135)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="Factor Xa site"
                     /translation="IEGR"
     polyA_signal    2280..2487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2496..2625
                     /label=R-ITR
                     /note="R-ITR"
     rep_origin      2734..3189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication;
                     arrowindicates direction of (+) strand synthesis"
     promoter        3521..3625
                     /gene="bla"
                     /label=AmpR promoter
                     /note="bla promoter"
     CDS             3626..4486
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4657..5245
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
       61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcacgcg ttctagaaat attaaggtac gggaggtact tggagcggcc
      241 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      301 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      361 tccccagtgc aagtgcaggt gccagaacat ttctctattc tagactcgag ctgcagaggg
      421 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      481 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      541 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      601 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      661 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      721 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      781 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      841 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgaattcgcc accatgtcta
      901 gactggacaa gagcaaagtc ataaactctg ctctggaatt actcaatgga gtcggtatcg
      961 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact
     1021 ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc
     1081 atcataccca ctcctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg
     1141 ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca
     1201 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc
     1261 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg
     1321 gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca
     1381 ccgattctat gcccccactt ctgaaacaag caattgagct gttcgaccgg cagggagccg
     1441 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt
     1501 gcgaaagcgg cgggccgacc gacgcccttg acgattttga cttagacatg ctcccagccg
     1561 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc
     1621 ttgacatgct ccccgggtaa aagcttatcg ataatcaacc tctggattac aaaatttgtg
     1681 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1741 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1801 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1861 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1921 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1981 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2041 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2101 ggacgccctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2161 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2221 ccctttgggc cgcctccccg catcgatacc gtcgactaga gctcgctgat cagcctcgac
     2281 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2341 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2401 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2461 ggaagacaat agcaggcatg ctggggagag atctgaggaa cccctagtga tggagttggc
     2521 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2581 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gagtggcccc
     2641 cccccccccc ccccccctgc agcctggcgt aatagcgaag aggcccgcac cgatcgccct
     2701 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     2761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3181 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3241 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3421 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3481 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3841 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4141 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4321 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4801 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5221 gtcagggggg cggagcctat ggaaaaacg
//
