LOCUS       Exported                7660 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1 - HA-KLF4 FL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7660)
  AUTHORS   Lin CC, Liu LZ, Addison JB, Wonderlin WF, Ivanov AV, Ruppert JM
  TITLE     A KLF4-miRNA-206 autoregulatory feedback loop can promote or inhibit
            protein translation depending upon cell context.
  JOURNAL   Mol Cell Biol. 2011 Jun;31(12):2513-27. Epub 2011 Apr 25.
  PUBMED    21518959
REFERENCE   2  (bases 1 to 7660)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7660
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             952..978
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3253..3270)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3259..3483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3529..3957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3616..3635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3826..3847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3966..3986)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3971..4300
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4151..4286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4213..4232
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4367..5161
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4421..4440)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5031..5050
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5335..5456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5372..5391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5426..5445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5505..5521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5505..5521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5518..5540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5529..5545
                     /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(5553..5583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5598..5619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5736..5753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5907..6495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5987..6006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6666..7526)
                     /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"
     primer_bind     7289..7308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7527..7631)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgg taccgagctc ggatccggcg agtctgacat gtacccctac
      961 gacgtgcccg actacgccgc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg
     1021 tctggcccgg cgggaagaga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg
     1081 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac
     1141 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc
     1201 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac
     1261 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg
     1321 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg
     1381 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg
     1441 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc
     1501 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg
     1561 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg
     1621 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc
     1681 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg
     1741 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg
     1801 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac
     1861 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg
     1921 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg
     1981 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat
     2041 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga
     2101 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc
     2161 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa
     2221 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc
     2281 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca
     2341 ttttccaggt cggaccacct cgccttacac atgaagaggc atttttaaat cccagacagt
     2401 ggatatgacc cacactgcca gaagagaatt cagtattttt tacttttcac actgtcttcc
     2461 cgatgaggga aggagcccag ccagaaagca ctacaatcat ggtcaagttc ccaactgagt
     2521 catcttgtga gtggataatc aggaaaaatg aggaatccaa aagacaaaaa tcaaagaaca
     2581 gatggggtct gtgactggat cttctatcat tccaattcta aatccgactt gaatattcct
     2641 ggacttacaa aatgccaagg gggtgactgg aagttgtgga tatcagggta taaattatat
     2701 ccgtgagttg ggggagggaa gaccagaatt cccttgaatt gtgtattgat gcaatataag
     2761 cataaaagat caccttgtat tctctttacc ttctaaaagc cattattatg atgttagaag
     2821 aagaggaaga aattcaggta cagaaaacat gtttaaatag cctaaatgat ggtgcttggt
     2881 gagtcttggt tctaaaggta ccaaacaagg aagccaaagt tttcaaactg ctgcatactt
     2941 tgacaaggaa aatctatatt tgtcttccga tcaacattta tgacctaagt caggtaatat
     3001 acctggttta cttctttagc atttttatgc agacagtctg ttatgcactg tggtttcaga
     3061 tgtgcaataa tttgtacaat ggtttattcc caagtatgcc ttaagcagaa caaatgtgtt
     3121 tttctatata gttccttgcc ttaataaata tgtaatataa atttaaaaaa aaaaaaaaaa
     3181 aaaactcgac ctgagtaact gacacaatgg atcgggtcga gtctagaggg cccgtttaaa
     3241 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3301 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3361 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3421 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3481 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3541 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3601 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3661 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3721 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3781 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3841 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3901 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3961 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4021 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4141 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4201 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4261 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4321 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4381 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4441 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4501 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4561 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4621 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4681 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4741 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4801 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4861 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4921 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4981 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5041 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5101 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5161 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5221 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5281 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5341 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5401 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5461 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5521 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5581 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5641 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5701 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5761 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5821 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5881 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5941 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6001 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6061 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6121 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6181 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6241 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6301 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6361 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6421 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6481 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6541 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6601 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6661 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6721 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6781 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6841 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6901 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6961 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7021 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7081 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7141 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7201 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7261 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7321 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7381 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7441 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7501 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7561 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7621 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
//
