LOCUS       sequence-14397-e        8129 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 8129)
  AUTHORS   Wirsing A, Senkel S, Klein-Hitpass L, Ryffel GU
  TITLE     A systematic analysis of the 3'UTR of HNF4A mRNA reveals an 
            interplay of regulatory elements including miRNA target sites.
  JOURNAL   PLoS One. 2011;6(11):e27438. Epub 2011 Nov 30.
  PUBMED    22140441
REFERENCE   2  (bases 1 to 8129)
  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: "PLoS One.";
            date: "2011"; volume: "6(11)"; pages: "e27438. Epub 2011 Nov 30"
COMMENT     Sequence Label: pRLCon1-1746
FEATURES             Location/Qualifiers
     source          1..8129
                     /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    3833..3954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3844..3863)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3898..3917
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4140..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4712..5069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4712..4903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4735..4755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4920..5055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5120..5914
                     /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(5174..5193)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5784..5803
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5978..6026
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6067..6086)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6186..6208
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6246..6264)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6332..6436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6437..6505,6506..7297)
                     /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(6655..6674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7468..8056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7957..7976
                     /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 ttctagttag
     2041 caagccgctg gggcttgggg gctccactgg ctccccccag ccccctaaga gagcacctgg
     2101 tgatcacgtg gtcacggcaa aggaagacgt gatgccagga ccagtcccag agcaggaatg
     2161 ggaaggatga agggcccgag aacatggcct aagggccaca tcccactgcc acccttgacg
     2221 ccctgctctg gataacaaga ctttgacttg gggagacctc tactgccttg gacaactttt
     2281 ctcatgttga agccactgcc ttcaccttca ccttcatcca tgtccaaccc ccgacttcat
     2341 cccaaaggac agccgcctgg agatgacttg aggccttact taaacccagc tcccttcttc
     2401 cctagcctgg tgcttctcct ctcctagccc ctgtcatggt gtccagacag agccctgtga
     2461 ggctgggtcc aattgtggca cttggggcac cttgctcctc cttctgctgc tgcccccacc
     2521 tctgctgcct ccctctgctg tcaccttgct cagccatccc gtcttctcca acaccacctc
     2581 tccagaggcc aaggaggcct tggaaacgat tcccccagtc attctgggaa catgttgtaa
     2641 gcactgactg ggaccaggca ccaggcaggg tctagaaggc tgtggtgagg gaagacgcct
     2701 ttctcctcca acccaacctc atcctccttc ttcagggact tgggtgggta cttgggtgag
     2761 gatccctgaa ggccttcaac ccgagaaaac aaacccaggt tggcgactgc aacaggaact
     2821 tggagtggag aggaaaagca tcagaaagag gcagaccatc caccaggcct ttgagaaagg
     2881 gtagaattct ggctggtaga gcaggtgaga tgggacattc caaagaacag cctgagccaa
     2941 ggcctagtgg tagtaagaat ctagcaagaa ttgaggaaga atggtgtggg agagggatga
     3001 tgaagagaga gagggcctgc tggagagcat agggtctgga acaccaggct gaggtcctga
     3061 tcagcttcaa ggagtatgca gggagctggg cttccagaaa atgaacacag cagttctgca
     3121 gaggacggga ggctggaagc tgggaggtca ggtggggtgg atgatataat gcgggtgaga
     3181 gtaatgaggc ttggggctgg agaggacaag atgggtaaac cctcacatca gagtgacatc
     3241 caggaggaat aagctcccag ggcctgtctc aagctcttcc ttactcccag gcactgtctt
     3301 aaggcatctg acatgcatca tctcatttaa tcctcccttc ctccctatta acctagagat
     3361 tgtttttgtt ttttattctc ctcctccctc cccgccctca cccgccccac tccctcctaa
     3421 cctagagatt gttacagaag ctgaaattgc gttctaagag gtgaagtgat tttttttctg
     3481 aaactcacac aactaggaag tggctgagtc aggacttgaa cccaggtctc cctggatcag
     3541 aacaggagct cttaactaca gtggctgaat agcttctcca aaggctccct gtgttctcac
     3601 cgtgatcaag ttgaggggct tccggctccc ttctacagcc tcagaaacca gactcgttct
     3661 tctgggaacc ctgcccactc ccaggaccaa gattggcctg aggctgcact aaaattcact
     3721 tagggtcgag catcctgttt gctgataaat attaaggaga attcatgact cttgacagct
     3781 tttctcgcgg ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac
     3841 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3901 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3961 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4021 aagtaaaacc tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata
     4081 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     4621 catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa
     4681 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     4741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4981 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5041 gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga
     5101 acttaaggct agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5161 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5221 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5281 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5341 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5401 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5461 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5521 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5581 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5641 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5701 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5761 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5821 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5881 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5941 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     6001 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac
     6061 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6121 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6181 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     6241 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6301 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6361 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6421 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6481 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6541 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6601 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6661 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6721 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6781 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6841 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6901 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6961 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7021 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7081 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7141 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7201 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7261 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8101 gccttttgct cacatggctc gacagatct
//
