LOCUS       Exported                5647 bp ds-DNA     circular SYN 05-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5647)
  AUTHORS   Path-IT
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 1, 2020 from SnapGene 5.1.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5647
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             1768..1791
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1828..1851
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1858..1881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1894..1916
                     /label=PreScission site
                     /note="SerialCloner_Color="
     primer_bind     1903..1949
                     /label=ULK1MTIsgibfor
     primer_bind     1924..1944
                     /label=ULK1_832_Bsa1_for
     primer_bind     2344..2362
                     /label=ULK1_MIT2_start
     primer_bind     complement(2555..2604)
                     /label=ULK1MITsGibrev
     polyA_site      2734..2789
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3150..3166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3174..3190
                     /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)."
     promoter        complement(3198..3228)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3243..3264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3323..3518
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3369..3504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3524..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3897..4485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4656..5516)
                     /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(5517..5621)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgatc
     1741 gacggtatcg atgccaccat ggctagctgg agccacccgc agttcgagaa aggtggaggt
     1801 gcccgaggtg gatcgggagg tggatcgtgg agccacccgc agttcgaaaa aggattcgat
     1861 tacaaggatg acgatgacaa gggcacaaga tctctggaag tcctgttcca gggcccgggt
     1921 accgagaccc tcatggagca agagcacacg gagatcctgc gtggcctgcg cttcacgctg
     1981 ctgttcgtgc agcacgtcct ggagatcgca gccctgaagg gcagcgccag tgaggcggcg
     2041 gggggccctg agtaccagct gcaggagagt gtggtggccg accagatcag cctgctgagc
     2101 cgagaatggg gcttcgcgga acagctggtg ctgtacctga aggtggccga gctactgtcc
     2161 tccggcctgc aaagtgccat cgaccagatc cgggccggca agctctgcct gtcgtccact
     2221 gtgaagcagg tggtgcgcag gctgaatgag ctgtacaagg ccagcgtggt gtcctgccag
     2281 ggcctgagcc tgcggctgca gcgcttcttc ctggacaagc agcggctcct ggaccgcatt
     2341 cacagcatca ctgccgagag gctcatcttc agccacgctg tgcagatggt gcagtcggct
     2401 gccctggacg agatgttcca gcaccgtgag ggctgcgtcc cacgctacca caaggccctg
     2461 ctgctcctgg aggggctgca gcacatgctc tcggaccagg ccgacatcga gaacgtcacc
     2521 aagtgcaagc tgtgcattga gcggagactc tcggcgctgc tgactggcat ctgtgcctga
     2581 tcgaggaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     2641 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     2701 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     2761 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     2821 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     2881 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     2941 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     3001 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     3061 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     3121 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3181 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3241 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3301 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     3361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3421 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3481 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     3541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     3661 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3781 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4081 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4261 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4321 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5641 ccacctg
//
