LOCUS       sequence-468440-        9141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the 3' UTR in regulating HRH1-P2 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9141)
  TITLE     Study the role and mechanism of the 3' UTR in regulating gene 
            function.
REFERENCE   2  (bases 1 to 9141)
  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"
COMMENT     Sequence Label: HRH1-P2
FEATURES             Location/Qualifiers
     source          1..9141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(747..1538,1539..1607))
                     /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     1370..1389
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1608..1712)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    1868..1916
                     /note="synthetic polyadenylation signal"
     CDS             complement(1980..2771)
                     /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="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2091..2110)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2701..2720
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2823..3755)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     regulatory      3752..3761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3806..4163)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3820..3955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3972..4163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4120..4140
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4408..4529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4445..4464)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4499..4518
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6488..8140)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     regulatory      8137..8146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8187..8686)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8235..8254)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8628..8651
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    8743..9026
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaattac caatgcttaa tcagtgaggc acctatctca
      781 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      841 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      901 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      961 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1021 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1081 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1141 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat agttgtcaga
     1201 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1261 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1321 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1381 ccacatagca gaactttgaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1441 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1501 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1561 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1621 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1681 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1741 gtctaagaaa ccattacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     1801 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagaggat ccttatcgct
     1861 atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     1921 ccatcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     1981 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     2041 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     2101 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     2161 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     2221 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     2281 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     2341 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     2401 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     2461 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     2521 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     2581 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     2641 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     2701 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     2761 tcttgttcaa tccctccgga gcctccgcca cctgatccgc cacctccgct cccaccaccg
     2821 gtctgctcgt tcttcagcac gcgctccacg aagctcttga tgtacttacc catttcatct
     2881 ggagcgtcct cctggctgaa gtggaggccc ttcaccttca cgaactcggt gttagggaac
     2941 ttcttagctc cctcgacaat agcgttggaa aagaacccag ggtcggactc gatgaacatc
     3001 ttaggcagat cgtcgctggc ccgaaggtag gcgttgtagt tgcggacaat ctggacgacg
     3061 tcgggcttgc ctcccttaac gagagggatc tcgcgaggcc aggagagggt aggccgtcta
     3121 acctcgccct tctccttgaa tggctccagg taggcagcga actcctcagg ctccagtttc
     3181 cgcatgatct tgcttgggag catggtctcg acgaagaagt tattctcaag caccattttc
     3241 tcgccctctt cgctcttgat cagggcgata tcctcctcga tgtcaggcca ctcgtcccag
     3301 gactcgatca cgtccacgac actctcagca tggacgatgg ccttgatctt gtcttggtgc
     3361 tcgtaggagt agtgaaaggc cagacaagcc ccccagtcgt ggcccacaaa gatgattttc
     3421 tttggaaggt tcagcagctc gaaccaagcg gtgaggtact tgtagtgatc caggaggcga
     3481 tatgagccat tcccgctctt gccggactta cccattccga tcagatcagg gatgatgcat
     3541 ctagccacgg gctcgatgtg aggcacgacg tgcctccaca ggtagctgga ggcagcgtta
     3601 ccatgcagaa aaatcacggc gttctcggcg tgcttctcgg aatcatagta gttgatgaag
     3661 gagtccagca cgttcatttg cttgcagcga gcccaccact gaggcccagt gatcatgcgt
     3721 ttgcgttgct cggggtcgta caccttggaa gccatggtgg ctccagcctt tggttcgaga
     3781 ctgttgtgtc agaagaatca agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     3841 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     3901 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     3961 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     4021 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     4081 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     4141 acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt
     4201 caggccatgg tgctgcgcag atccgcgtat gcggtgtgaa ataccgcaca gatgcgtaag
     4261 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgattttacc
     4321 acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct gaacctgaaa
     4381 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4441 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4501 ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc gccccaaggg
     4561 gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt
     4621 tagcagccgg atcagcttgc atgcctgcag gtcgactagt atgcttttaa acatcatatt
     4681 ttaggcaggt aaaaatatct cactttgaca gatcatgcaa agaggaacac aaaatacaaa
     4741 acatcagctt gtacaggcac tccatatgct atacatattg tggtttagca ccaagcttta
     4801 aaatataaat cacaaccaga ttgcaacata aacaccaagt aaaactacat gttcctatgt
     4861 ctttactagt ttttgcttac caggtatggc tatgaaagat tacaatgact ttttgacaaa
     4921 cacaacagaa ctcacagata aaaatgcact tggttccttc aaagcgtccc cttccgtagg
     4981 tgcacgggaa ctccatgtca gctgttcgtc tactcaaaca tatcattttg agctgccacc
     5041 actcttggga ataccgtgta tgtgtatctg tgcacactta agagggcaca ttgttaaaat
     5101 attgtatttt tttttttgag acagagtctt gctctgttgc ccaggctgga gtgcagtggc
     5161 gtgatcttgg ctcacggcaa cctccacctc ccgggttcaa gcaattctcg tgcctcggcc
     5221 tcccaagtaa gtgggactac aggcatgccc caccatgccc agataatttt tgtgttttta
     5281 gtagagacaa ggtttctcca tattggccag actggtcttg aactcctggc ctcaaatgat
     5341 ctgcccacct cagcctctca acatgctggg attataggct tgagctacca tgcccggcca
     5401 aaagggcaca tttttgcctt tcggagaaaa tatgctacaa taagctgaaa ccccagagga
     5461 tctgtactct aactttctca cataactagc tatgtgacct tgggtacctc tctcaccttg
     5521 ctgaacctca atttgctcct ctgcaaacta gagttagaaa attaccctcc tgtggggtgt
     5581 tgtggggctt aaatgagata aacctagtac agccctcaaa aatatggcat tctgccccca
     5641 ttccttcttt taatttttaa cgtgcaacca ttcaaaaata tttcatttct ctttcttttg
     5701 aggatgtctt ttgccaccac ttttttaata gccaaaggaa atttggggtt aaagacgatg
     5761 ggtgtgcaag ctgcataatt ttctatccac tctaaacatg tttgagtaag aaataataag
     5821 gcagaatttc cagggtggtt cagacactgg ttctggagaa agctgttaaa gaggctcaga
     5881 agagaagttt tctgagaagg gatagcccct gtggaacagc tgtcatgcct ctcaaggtga
     5941 cccctggaag gggacagcaa ctattggcat ggataaagtc tctgcacagt tatacagctc
     6001 cattccagga gggctcaaga gtccttttca attctccagt ggaaccctag ccacgtgtct
     6061 cttcagttgt caatcacttg actcggaaag cttgaattac aatgagtctg aggctcccat
     6121 agttcctgag gagcaggaga gttccaccca ggtctgattc tctctctctt ttatttttat
     6181 tatttttttc tttttgaacc cagttacaag aaacaggtct gactttcttg caaagattct
     6241 gcttcctcct caaagttcaa tctctctgct gatcaccgcc atctgctgtt cttctatggt
     6301 gcctaagaac tttccaaact accaagcccc taagactgtg gtttggattc cagaaagccc
     6361 aggtgcctgc ctggcaacac acaggccttc gtcctctatt tccttgttgg acatcataag
     6421 gatcattttg ttgcatcccc tcagagcctc ccactcgagg ctagcgagct cgtttaaaca
     6481 actagaatta cacggcgatc ttgccgccct tcttggcctt aatgagaatc tcgcggatct
     6541 tgcgggcgtc caacttgccg gtcagtcctt taggcacctc gtccacgaac acaacaccac
     6601 cgcgcagctt cttggcggtt gtaacctggc tggccacata gtccacgatc tccttctcgg
     6661 tcatggtttt accgtgttcc agcacgacga ctgcggcggg cagctcgccg gcatcgtcgt
     6721 cgggcaggcc ggcgaccccg gcgtcgaaga tgttggggtg ttgcagcagg atgctctcca
     6781 gttcggctgg ggctacctgg tagcccttgt atttgatcag gctcttcagc cggtccacga
     6841 tgaagaagtg ctcgtcctcg tcccagtagg cgatgtcgcc gctgtgcagc cagccgtcct
     6901 tgtcgatgag agcgtttgta gcctcggggt tgttaacgta gccgctcatg atcatggggc
     6961 cacggacgca cagctcgccg cgctggttca cacccagtgt cttaccggtg tccaagtcca
     7021 ccaccttagc ctcgaagaag ggcaccacct tgcctactgc gccaggcttg tcgtcccctt
     7081 cgggggtgat cagaatggcg ctggttgttt ctgtcaggcc gtagccctgg cggatgcctg
     7141 gtaggtggaa gcgtttggcc acggcctcac ctacctcctt gctgagcggc gccccgccgc
     7201 tggcgatctc gtgcaagttg cttaggtcgt acttgtcgat gagagtgctc ttagcgaaga
     7261 agctaaatag tgtgggcacc agcagggcag attgaatctt atagtcttgc aagctgcgca
     7321 agaatagctc ctcctcgaag cggtacatga gcacgacccg aaagccgcag atcaagtagc
     7381 ccagcgtggt gaacatgccg aagccgtggt gaaatggcac cacgctgagg atagcggtgt
     7441 cggggatgat ctggttgccg aagatggggt cgcgggcatg actgaatcgg acacaagcgg
     7501 tgcggtgcgg tagggctacg cccttgggca atccggtact gccactactg ttcatgatca
     7561 gggcgatggt tttgtcccgg tcgaagctct cgggcacgaa gtcgtactcg ttgaagccgg
     7621 gtggcaaatg ggaagtcacg aaggtgtaca tgctttggaa gccctggtag tcggtcttgc
     7681 tatccatgat gatgatcttt tgtatgatcg gtagcttctt ttgcacgttg aggatctttt
     7741 gcagcccttt cttgctcacg aatacgacgg tgggctggct gatgcccatg ctgttcagca
     7801 gctcgcgctc gttgtagatg tcgttagctg gggccacagc cacaccgatg aacagggcac
     7861 ccaacacggg catgaagaac tgcaagctat tctcgctgca caccacgatc cgatggtttg
     7921 tattcagccc atagcgcttc atagcttctg ccagccgaac gctcatctcg aagtactcgg
     7981 cgtaggtaat gtccacctcg atatgtgcgt cggtaaaggc gatggtgccg ggcaccaggg
     8041 cgtagcgctt catggctttg tgcagctgct cgccggcggt cccgtcttcg agtgggtaga
     8101 atggcgctgg gcccttctta atgtttttgg catcttccat ggtggcttta ccaacagtac
     8161 cggattgcca agcttgggct gcaggtcgaa aggcccggag atgaggaaga ggagaacagc
     8221 gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga ccttcgggcg
     8281 cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct tcccagcctc
     8341 tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag gcctacccgc
     8401 ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc tacttccatt
     8461 tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac taggggagga
     8521 gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct accggtggat
     8581 gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag cggggctgct
     8641 aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggta gatctcgatc
     8701 ctctacgccg gacgcatcgt ggccggagta ctgcggttta tctcgatggc tacgagggca
     8761 gacagtaagt ggatttacca taatccctta attgtacgca ccgctaaaac gcgttcagcg
     8821 cgatcacggc agcagacagg taaaaatggc aacaaaccac ccgaaaaact gccgcaatcg
     8881 cgcctgataa attttaaccg tatgaatacc tatgcaacca gagggtacag gccacattac
     8941 ccccacttaa tccactgaag ctgccatttt tcatggtttc accatcccag cgaagggcca
     9001 tccagcgtgc gttcctgtat ttccggacta taaaaggatc taggtgaaga tcctttttga
     9061 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     9121 agaaaagatc aaaggatctt c
//
