LOCUS       sequence-14404-e        6632 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 6632)
  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 6632)
  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: pRLCon1-249
FEATURES             Location/Qualifiers
     source          1..6632
                     /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    2336..2457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2347..2366)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2401..2420
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2643..3098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2730..2749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2940..2961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3215..3572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3215..3406
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3238..3258)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3423..3558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3623..4417
                     /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(3677..3696)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4287..4306
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4481..4529
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4570..4589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4689..4711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4749..4767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4835..4939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4940..5008,5009..5800)
                     /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(5158..5177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5971..6559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6460..6479
                     /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 ctcatgttgg cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga
     2341 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2401 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2461 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     2521 agcaagtaaa acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta
     2581 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2641 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2701 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2761 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2821 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2881 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2941 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3001 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3061 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3121 acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa
     3181 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     3241 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3301 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3421 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3481 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3541 gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct
     3601 cgaacttaag gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc
     3661 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3721 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3781 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3841 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3901 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3961 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4021 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4081 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4141 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4201 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4261 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4321 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4381 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4441 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     4501 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg
     4561 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4621 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4681 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4741 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4801 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4921 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4981 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5041 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5101 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5161 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5221 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5281 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5341 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5401 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5461 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5521 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5581 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5641 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5701 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5761 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6601 ctggcctttt gctcacatgg ctcgacagat ct
//
