LOCUS       sequence-44813-e        8144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  AUTHORS   Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, 
            Vadas MA, Khew-Goodall Y, Goodall GJ
  TITLE     The miR-200 family and miR-205 regulate epithelial to mesenchymal 
            transition by targeting ZEB1 and SIP1.
  JOURNAL   Nat Cell Biol. 2008 May;10(5):593-601. Epub 2008 Mar 30.
  PUBMED    18376396
REFERENCE   2  (bases 1 to 8144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol."; date: "2008-05"; volume: "10(5)"; pages: "593-601. Epub 2008
            Mar 30"
COMMENT     Sequence Label: pCI-neo-RL-ZEB1 200bmutx5
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..2030
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3848..3969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3859..3878)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3913..3932
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4155..4610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4242..4261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4452..4473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4727..5084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4727..4918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4750..4770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4935..5070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5135..5929
                     /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(5189..5208)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5799..5818
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5993..6041
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6082..6101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6201..6223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6261..6279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6347..6451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6452..6520,6521..7312)
                     /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     complement(6670..6689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7483..8071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7972..7991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggct tccaaggtgt acgaccccga gcaacgcaaa cgcatgatca
     1141 ctgggcctca gtggtgggct cgctgcaagc aaatgaacgt gctggactcc ttcatcaact
     1201 actatgattc cgagaagcac gccgagaacg ccgtgatttt tctgcatggt aacgctgcct
     1261 ccagctacct gtggaggcac gtcgtgcctc acatcgagcc cgtggctaga tgcatcatcc
     1321 ctgatctgat cggaatgggt aagtccggca agagcgggaa tggctcatat cgcctcctgg
     1381 atcactacaa gtacctcacc gcttggttcg agctgctgaa ccttccaaag aaaatcatct
     1441 ttgtgggcca cgactggggg gcttgtctgg cctttcacta ctcctacgag caccaagaca
     1501 agatcaaggc catcgtccat gctgagagtg tcgtggacgt gatcgagtcc tgggacgagt
     1561 ggcctgacat cgaggaggat atcgccctga tcaagagcga agagggcgag aaaatggtgc
     1621 ttgagaataa cttcttcgtc gagaccatgc tcccaagcaa gatcatgcgg aaactggagc
     1681 ctgaggagtt cgctgcctac ctggagccat tcaaggagaa gggcgaggtt agacggccta
     1741 ccctctcctg gcctcgcgag atccctctcg ttaagggagg caagcccgac gtcgtccaga
     1801 ttgtccgcaa ctacaacgcc taccttcggg ccagcgacga tctgcctaag atgttcatcg
     1861 agtccgaccc tgggttcttt tccaacgcta ttgtcgaggg agctaagaag ttccctaaca
     1921 ccgagttcgt gaaggtgaag ggcctccact tcagccagga ggacgctcca gatgaaatgg
     1981 gtaagtacat caagagcttc gtggagcgcg tgctgaagaa cgagcagtaa ttctagtcaa
     2041 aataaatccg ggtgtgcctg aacctcagac ctagtaattt ttcatgcagt tttcaaagtt
     2101 aggaacaagt ttgtaacatg cagcagatta gaaaacctta atgactcaga gagcaacaat
     2161 acaagaggtt aaaggaagct gattaattag atatgcatct ggcattgttt tatcttatca
     2221 ggactatcac tcttatgttg gtttattctt aagctgtaca attgggagaa attttataat
     2281 tttttattgg taaacatatg ctaaatccgc ttcaggactt tattatgttt tttaaaatgt
     2341 gagaacttct gcactacaaa attcccttca cagagaagta taatgtagtt ccaacccgtg
     2401 ctaactacct tttataaatt cagtctagaa ggtagtaatt tctaatattt agatgtctta
     2461 gtagagcgta ttatcattta aagtgtattg ttagccttaa gaaagcagct gatagaagaa
     2521 ctgaagtttc ttactcacgt ggtttaaaat ggagttcaaa agattgccat tgagttctga
     2581 ttgcagggac taacaatgtt aatctgataa ggacagcaaa atcatcagaa tcagtgtttg
     2641 tgattgtgtt tgaatatgtg gtaacatatg aaggatatga catgaagctt tgtatctcct
     2701 ttggccttaa gcaagacctg tgtgctgtaa gtgccatttc tcaggacttt caaggctcta
     2761 acccgccttc atccaatgtg tggcctacaa taactagcat ttgttgattt gtctcttgta
     2821 tcaaaattcc caaataaaac ttaaaaccac tgactctgtc agagaaactg aaacactggg
     2881 acatttcatc cttcaattcc tcggtattga ttttatgttg attgattttc agaatttctc
     2941 tacagaaacg aaagggaaat tttctaatct gctttatcca tgtacttgca tttcagacat
     3001 ggacatgcta ttgttatttg gctcataact gtttccaaat gttagttatt atggacccaa
     3061 tttattaaca acattagctg atttttacct atcaggacta ttttatttct tttagtttat
     3121 agatctgtgc aacatttttg tactgtatgt cttcaaacct ggcaggacta atacccttct
     3181 tactgacata tgtactttta gttttagaaa acttttatat ttatgtgtct tatttttata
     3241 tttctttatt tattacacag tgtagtgtat aatactgtag tttgtattaa tacaataata
     3301 tattttagta tgaaaatttg gaaagttgat aagatttaaa gtagagatgc aattggttct
     3361 cctgcattga gatttgattt aacagtgtta tgttaacatt tatacttgcc ttggactgta
     3421 gaacagaact taaatgggaa tgtattagtt ttacaactac aatcaagtca ttttaccttt
     3481 acccagtttt taatataaaa cttaaatttt gaaattcact gtgtgactaa tagcatgatg
     3541 ctctgcagtt ttattaagaa atcagcctaa ccatacaact ctcatttcct tagtaagcca
     3601 aattaggatt aacttctata aacagtgttg ggaacaatgt ttaacatttt gtgccaattt
     3661 gttcctgtat tcatgtatgt aagttacaga tctgactctt catttttaag ttccttgtta
     3721 catcatggtc attttctagt tttttaccag actcccatct cacaataaaa tgcatcaaca
     3781 agcctgaact gctgtactag agcggccgct tccctttagt gagggttaat gcttcgagca
     3841 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3901 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3961 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     4021 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatccgata aggatcgatc
     4081 cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4141 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4201 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4501 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4561 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     4621 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac
     4681 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     4741 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg
     5101 acacaacagt ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc
     5161 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5221 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5281 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5341 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5401 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5461 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5521 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5581 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5641 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5701 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5761 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5821 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5881 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5941 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     6001 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     6061 ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6121 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6181 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6241 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6301 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6421 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6481 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6541 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6601 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6661 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6721 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6781 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6841 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6901 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6961 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7021 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7081 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7141 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7201 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7261 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7321 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7381 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7501 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7561 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7621 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7681 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7741 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7801 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7861 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7921 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7981 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8041 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8101 cctggccttt tgctggcctt ttgctcacat ggctcgacag atct
//
