LOCUS       sequence-472284-        8889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the P4HA1-P2 3' UTR in regulation 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8889)
  TITLE     Study the role and mechanism of the 3' UTR in regulating gene 
            function.
REFERENCE   2  (bases 1 to 8889)
  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: P4HA1-P2
FEATURES             Location/Qualifiers
     source          1..8889
                     /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(6236..7888)
                     /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      7885..7894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7935..8434)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7983..8002)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8376..8399
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    8491..8774
                     /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 gtcgactcaa acaggcttcc ctttttctcc
     4681 tattgttgta ctcttatgtg tctgatatac acatttccta gtcttaactt tcaggagttt
     4741 acaattgact aacactccat gattgattca gtcatgaacc tcatcccatg tttcatctgt
     4801 ggacaattgc ttactttgtg ggttctttta aaagtaacac gaaatcatca tattgcataa
     4861 aaccttaaag ttctgttggt atcacagaag acaaggcaga gtttaaagtg aggaatttta
     4921 tatttaaaga actttttggt tggataaaaa cataatttga gcatccagtt ttagtatttc
     4981 actacatctc agttggtggg tgttaagcta gaatgggctg tgtgatagga aacaaatgcc
     5041 ttacagatgt gcctaggtgt tctgtttacc tagtgtctta ctctgttttc tggatctgaa
     5101 gactagtaat aaactaggac actaactggg ttccatgtga ttgccctttc atatgatctt
     5161 ctaagttgat ttttttcctc ccaagtcttt tttaaagaaa gtatactgta ttttaccaac
     5221 cccctctctt ttcttttagc tcctctgtgg tgaattaaac gtacttgagt taaaatattt
     5281 cgattttttt tttttaatgg aaagtcctgc ataacaacac tgggccttct taactaaaat
     5341 gctcaccact tagcctgttt ttttatccct tttttaaaat gacagatgat tttgttcagg
     5401 aattttgctg tttttcttag tgctaatacc ttgcctctta ttcctgctac agcagggtgg
     5461 taatattggc attctgatta aatactgtgc cttaggagac tggaagttta aaaatgtaca
     5521 agtcctttca gtgatgaggg aattgatttt ttttaaaagt ctttttctta gaaagccaaa
     5581 atgtttgttt ttttaagatt ctgaaatgtg ttgtgacaac aatgacctat ttatgatctt
     5641 aaatcttttt taaaaaaatg tttgttttct gtgtggtgtt tttcgtattt aaatccgaat
     5701 gtatgatgtg gcagtaacag gttaacttat gtaatttctt tagtacatag ggcttaggtt
     5761 tatactcttg gtttccactc acactaatgt cacatggtta agaaaaactt aggctctctg
     5821 aagtttcaga tatttcatga atcaagatgt gcttccttca aagagagctc taaaatactt
     5881 tactgtggtt atgaagttat tatgggccag gcatggtggc tcacgccttc aatcccagca
     5941 ctttgggagg ccgaggcagg cggatcacct gaggctagga gttcgagacc agcctggcca
     6001 acatggtgaa accccatccc tactaaaatt accaaaatta gccgggtgtg gtggtgcatg
     6061 cctgtagtcc cagctacttg ggaggctgag gcaggaaaat cgcttgaacc tgggaggcag
     6121 aggttgcagt gagctgagat tgcaccactg cactctagct tgagtgacag agtgactctg
     6181 tctcaaaaaa gaaaaaaatt actcgaggct agcgagctcg tttaaacaac tagaattaca
     6241 cggcgatctt gccgcccttc ttggccttaa tgagaatctc gcggatcttg cgggcgtcca
     6301 acttgccggt cagtccttta ggcacctcgt ccacgaacac aacaccaccg cgcagcttct
     6361 tggcggttgt aacctggctg gccacatagt ccacgatctc cttctcggtc atggttttac
     6421 cgtgttccag cacgacgact gcggcgggca gctcgccggc atcgtcgtcg ggcaggccgg
     6481 cgaccccggc gtcgaagatg ttggggtgtt gcagcaggat gctctccagt tcggctgggg
     6541 ctacctggta gcccttgtat ttgatcaggc tcttcagccg gtccacgatg aagaagtgct
     6601 cgtcctcgtc ccagtaggcg atgtcgccgc tgtgcagcca gccgtccttg tcgatgagag
     6661 cgtttgtagc ctcggggttg ttaacgtagc cgctcatgat catggggcca cggacgcaca
     6721 gctcgccgcg ctggttcaca cccagtgtct taccggtgtc caagtccacc accttagcct
     6781 cgaagaaggg caccaccttg cctactgcgc caggcttgtc gtccccttcg ggggtgatca
     6841 gaatggcgct ggttgtttct gtcaggccgt agccctggcg gatgcctggt aggtggaagc
     6901 gtttggccac ggcctcacct acctccttgc tgagcggcgc cccgccgctg gcgatctcgt
     6961 gcaagttgct taggtcgtac ttgtcgatga gagtgctctt agcgaagaag ctaaatagtg
     7021 tgggcaccag cagggcagat tgaatcttat agtcttgcaa gctgcgcaag aatagctcct
     7081 cctcgaagcg gtacatgagc acgacccgaa agccgcagat caagtagccc agcgtggtga
     7141 acatgccgaa gccgtggtga aatggcacca cgctgaggat agcggtgtcg gggatgatct
     7201 ggttgccgaa gatggggtcg cgggcatgac tgaatcggac acaagcggtg cggtgcggta
     7261 gggctacgcc cttgggcaat ccggtactgc cactactgtt catgatcagg gcgatggttt
     7321 tgtcccggtc gaagctctcg ggcacgaagt cgtactcgtt gaagccgggt ggcaaatggg
     7381 aagtcacgaa ggtgtacatg ctttggaagc cctggtagtc ggtcttgcta tccatgatga
     7441 tgatcttttg tatgatcggt agcttctttt gcacgttgag gatcttttgc agccctttct
     7501 tgctcacgaa tacgacggtg ggctggctga tgcccatgct gttcagcagc tcgcgctcgt
     7561 tgtagatgtc gttagctggg gccacagcca caccgatgaa cagggcaccc aacacgggca
     7621 tgaagaactg caagctattc tcgctgcaca ccacgatccg atggtttgta ttcagcccat
     7681 agcgcttcat agcttctgcc agccgaacgc tcatctcgaa gtactcggcg taggtaatgt
     7741 ccacctcgat atgtgcgtcg gtaaaggcga tggtgccggg caccagggcg tagcgcttca
     7801 tggctttgtg cagctgctcg ccggcggtcc cgtcttcgag tgggtagaat ggcgctgggc
     7861 ccttcttaat gtttttggca tcttccatgg tggctttacc aacagtaccg gattgccaag
     7921 cttgggctgc aggtcgaaag gcccggagat gaggaagagg agaacagcgc ggcagacgtg
     7981 cgcttttgaa gcgtgcagaa tgccgggcct ccggaggacc ttcgggcgcc cgccccgccc
     8041 ctgagcccgc ccctgagccc gcccccggac ccaccccttc ccagcctctg agcccagaaa
     8101 gcgaaggagc aaagctgcta ttggccgctg ccccaaaggc ctacccgctt ccattgctca
     8161 gcggtgctgt ccatctgcac gagactagtg agacgtgcta cttccatttg tcacgtcctg
     8221 cacgacgcga gctgcggggc gggggggaac ttcctgacta ggggaggagt agaaggtggc
     8281 gcgaaggggc caccaaagaa cggagccggt tggcgcctac cggtggatgt ggaatgtgtg
     8341 cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa agcgcatgct
     8401 ccagactgcc ttgggaaaag cgcctcccct acccggtaga tctcgatcct ctacgccgga
     8461 cgcatcgtgg ccggagtact gcggtttatc tcgatggcta cgagggcaga cagtaagtgg
     8521 atttaccata atcccttaat tgtacgcacc gctaaaacgc gttcagcgcg atcacggcag
     8581 cagacaggta aaaatggcaa caaaccaccc gaaaaactgc cgcaatcgcg cctgataaat
     8641 tttaaccgta tgaataccta tgcaaccaga gggtacaggc cacattaccc ccacttaatc
     8701 cactgaagct gccatttttc atggtttcac catcccagcg aagggccatc cagcgtgcgt
     8761 tcctgtattt ccggactata aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8821 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8881 aggatcttc
//
