LOCUS       sequence-469295-        8260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the NKIRAS2-P1 3' UTR in regulating 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8260)
  TITLE     Studying the role and mechanism of the 3' UTR in regulating gene 
            function. Batch 2.
REFERENCE   2  (bases 1 to 8260)
  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: NKIRAS2-P1
FEATURES             Location/Qualifiers
     source          1..8260
                     /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(5607..7259)
                     /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      7256..7265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7306..7805)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7354..7373)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7747..7770
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7862..8145
                     /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 gtcgactatt ggtaggggtg gggaattttt
     4681 tttttttttt cgagatggag tcttgctctg ttgaccaggc tggagtgcag tggcacgatc
     4741 tcggctcact gcagcctccg ccttctgagt tcaagcaatt ctcctgcctc agcctccccg
     4801 ggtagctggg attacaggca tgtgccacta cgcccggcta acttttgtat ttttggcaga
     4861 gatggggttt cacaatgttg gccaggctgg tcgtgaactc ctgacctcaa gtaatcgacc
     4921 cgccttggcc tcccaaagtg ctgggattac aggggtgagc cactgagccc agccaggggt
     4981 gggggattta aaacagaaaa ctacactccc aggaggccaa cttcttccag ccaagaaaag
     5041 gacaggagaa aggtcagggt gagggatgag gagacgccta atgccagtat aggagcatat
     5101 ttgtgtgtgc acaactaaat gcatacacac aatacctgga cacccttttt ctggagtcca
     5161 ggctggtgcc ataagtgtgt gtgtgtcgct gatgggaagc ccagagggtg gggcccctac
     5221 ctataatagg aactcagacc caagaaggga cactgctgcg tggtgactct gcttcctgtc
     5281 tggggcagct ggaagggaca gtgacccaca actgccccag gagacagtgg agcagatcct
     5341 ctaaaagctg ggatggggag taggctggga gggctgagga aggggatgtc aaccaagagt
     5401 atttgcactt cccagaggtg agagggagca aagtggctgg ggcgtggcct ggcggggagc
     5461 tccctggctg actgggaaag acagcttgtc ctctactttg ccctcaacac atctaccaga
     5521 gccccagaca ctgaaatggg gctgggatcg tgaaagagga acggcagctc tactcgaggc
     5581 tagcgagctc gtttaaacaa ctagaattac acggcgatct tgccgccctt cttggcctta
     5641 atgagaatct cgcggatctt gcgggcgtcc aacttgccgg tcagtccttt aggcacctcg
     5701 tccacgaaca caacaccacc gcgcagcttc ttggcggttg taacctggct ggccacatag
     5761 tccacgatct ccttctcggt catggtttta ccgtgttcca gcacgacgac tgcggcgggc
     5821 agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat gttggggtgt
     5881 tgcagcagga tgctctccag ttcggctggg gctacctggt agcccttgta tttgatcagg
     5941 ctcttcagcc ggtccacgat gaagaagtgc tcgtcctcgt cccagtaggc gatgtcgccg
     6001 ctgtgcagcc agccgtcctt gtcgatgaga gcgtttgtag cctcggggtt gttaacgtag
     6061 ccgctcatga tcatggggcc acggacgcac agctcgccgc gctggttcac acccagtgtc
     6121 ttaccggtgt ccaagtccac caccttagcc tcgaagaagg gcaccacctt gcctactgcg
     6181 ccaggcttgt cgtccccttc gggggtgatc agaatggcgc tggttgtttc tgtcaggccg
     6241 tagccctggc ggatgcctgg taggtggaag cgtttggcca cggcctcacc tacctccttg
     6301 ctgagcggcg ccccgccgct ggcgatctcg tgcaagttgc ttaggtcgta cttgtcgatg
     6361 agagtgctct tagcgaagaa gctaaatagt gtgggcacca gcagggcaga ttgaatctta
     6421 tagtcttgca agctgcgcaa gaatagctcc tcctcgaagc ggtacatgag cacgacccga
     6481 aagccgcaga tcaagtagcc cagcgtggtg aacatgccga agccgtggtg aaatggcacc
     6541 acgctgagga tagcggtgtc ggggatgatc tggttgccga agatggggtc gcgggcatga
     6601 ctgaatcgga cacaagcggt gcggtgcggt agggctacgc ccttgggcaa tccggtactg
     6661 ccactactgt tcatgatcag ggcgatggtt ttgtcccggt cgaagctctc gggcacgaag
     6721 tcgtactcgt tgaagccggg tggcaaatgg gaagtcacga aggtgtacat gctttggaag
     6781 ccctggtagt cggtcttgct atccatgatg atgatctttt gtatgatcgg tagcttcttt
     6841 tgcacgttga ggatcttttg cagccctttc ttgctcacga atacgacggt gggctggctg
     6901 atgcccatgc tgttcagcag ctcgcgctcg ttgtagatgt cgttagctgg ggccacagcc
     6961 acaccgatga acagggcacc caacacgggc atgaagaact gcaagctatt ctcgctgcac
     7021 accacgatcc gatggtttgt attcagccca tagcgcttca tagcttctgc cagccgaacg
     7081 ctcatctcga agtactcggc gtaggtaatg tccacctcga tatgtgcgtc ggtaaaggcg
     7141 atggtgccgg gcaccagggc gtagcgcttc atggctttgt gcagctgctc gccggcggtc
     7201 ccgtcttcga gtgggtagaa tggcgctggg cccttcttaa tgtttttggc atcttccatg
     7261 gtggctttac caacagtacc ggattgccaa gcttgggctg caggtcgaaa ggcccggaga
     7321 tgaggaagag gagaacagcg cggcagacgt gcgcttttga agcgtgcaga atgccgggcc
     7381 tccggaggac cttcgggcgc ccgccccgcc cctgagcccg cccctgagcc cgcccccgga
     7441 cccacccctt cccagcctct gagcccagaa agcgaaggag caaagctgct attggccgct
     7501 gccccaaagg cctacccgct tccattgctc agcggtgctg tccatctgca cgagactagt
     7561 gagacgtgct acttccattt gtcacgtcct gcacgacgcg agctgcgggg cgggggggaa
     7621 cttcctgact aggggaggag tagaaggtgg cgcgaagggg ccaccaaaga acggagccgg
     7681 ttggcgccta ccggtggatg tggaatgtgt gcgaggccag aggccacttg tgtagcgcca
     7741 agtgcccagc ggggctgcta aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc
     7801 tacccggtag atctcgatcc tctacgccgg acgcatcgtg gccggagtac tgcggtttat
     7861 ctcgatggct acgagggcag acagtaagtg gatttaccat aatcccttaa ttgtacgcac
     7921 cgctaaaacg cgttcagcgc gatcacggca gcagacaggt aaaaatggca acaaaccacc
     7981 cgaaaaactg ccgcaatcgc gcctgataaa ttttaaccgt atgaatacct atgcaaccag
     8041 agggtacagg ccacattacc cccacttaat ccactgaagc tgccattttt catggtttca
     8101 ccatcccagc gaagggccat ccagcgtgcg ttcctgtatt tccggactat aaaaggatct
     8161 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8221 actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
