LOCUS       sequence-14402-e        7195 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 7195)
  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 7195)
  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: pRLCon1-449_850-1207
FEATURES             Location/Qualifiers
     source          1..7195
                     /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    2899..3020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2910..2929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2964..2983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3206..3661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3778..4135
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3778..3969
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3801..3821)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3986..4121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4186..4980
                     /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(4240..4259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4850..4869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5044..5092
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5133..5152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5252..5274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5312..5330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5398..5502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5503..5571,5572..6363)
                     /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(5721..5740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6534..7122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7023..7042
                     /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 tctagttagc
     2041 aagccgctgg ggcttggggg ctccactggc tccccccagc cccctaagag agcacctggt
     2101 gatcacgtgg tcacggcaaa ggaagacgtg atgccaggac cagtcccaga gcaggaatgg
     2161 gaaggatgaa gggcccgaga acatggccta agggccacat cccactgcca cccttgacgc
     2221 cctgctctgg ataacaagac tttgacttgg ggagacctct actgccttgg acaacttttc
     2281 tcatgttgaa gccactgcct tcaccttcac cttcatccat gtccaacccc cgacttcatc
     2341 ccaaaggaca gccgcctgga gatgacttga ggccttactt aaacccagct cccttcttcc
     2401 ctagcctggt gcttctcctc tcctagcccc tgtcatggtg tccagacaga gccctgtgag
     2461 gctgggtcca attgtggcac ttggggcatc tagatggctg gtagagcagg tgagatggga
     2521 cattccaaag aacagcctga gccaaggcct agtggtagta agaatctagc aagaattgag
     2581 gaagaatggt gtgggagagg gatgatgaag agagagaggg cctgctggag agcatagggt
     2641 ctggaacacc aggctgaggt cctgatcagc ttcaaggagt atgcagggag ctgggcttcc
     2701 agaaaatgaa cacagcagtt ctgcagagga cgggaggctg gaagctggga ggtcaggtgg
     2761 ggtggatgat ataatgcggg tgagagtaat gaggcttggg gctggagagg acaagatggg
     2821 taaaccctca catcagagtg acatccagga gggcggccgc ttccctttag tgagggttaa
     2881 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2941 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3001 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3061 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     3121 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3181 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3661 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3721 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3781 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4021 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4081 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4141 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     4201 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4261 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4321 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4381 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4441 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4501 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4561 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4621 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4681 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4741 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4801 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4861 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4921 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4981 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     5041 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5101 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5161 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5221 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5281 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5341 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5401 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5461 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5521 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5581 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5641 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5701 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5761 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5821 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5881 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5941 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6001 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6061 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6361 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6661 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
