LOCUS       sequence-469286-        8179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the 3' UTR in regulating SPINT1-P1 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8179)
  TITLE     Studying the role and mechanism of the 3' UTR in regulating gene 
            function. Batch 3.
REFERENCE   2  (bases 1 to 8179)
  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: SPINT1-P1
FEATURES             Location/Qualifiers
     source          1..8179
                     /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(5526..7178)
                     /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      7175..7184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7225..7724)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7273..7292)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7666..7689
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7781..8064
                     /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 gtcgactctc tccaaagtca ggggccaggc
     4681 cagtttcgag agagacgatg ggaaaacctg ctcaacaggg tggctgaagc ctgaaccagc
     4741 tgaacaggac ggtcccatca ccccccagca acctagtccc tctcctggcc cacagaacag
     4801 agccagaaac tccacaaacc acttttatta cccagtgggc gggctgggct gtgatgttgg
     4861 agaaccttgg gggtgggggc tgcggaatgc agctgagctt ctcctggctc tgtctgctgg
     4921 tctaggccag ggtggggctg atcaagggca gagagctcaa tcttggggga agaggaagag
     4981 aggatagaga ggccaaacag gcctcttccc ctcctcttca cccatgccac agcattaaat
     5041 aaacaaaaag caactcttta cagcacaaac tacacaggga agtccttcct cccagccctg
     5101 ggcgcacagc atggaagagg agctcgggag gacagggttg gggcctccag ggtctgacac
     5161 caccccactc ctcttcctag caccatgggg tagggacagg cagctttgaa cagaggaaac
     5221 aaaactccaa cttcctccaa actcctatgc aggcaggtcc atctagtctg gtacttctgg
     5281 cccaagggtt ttctgctcct tccaaacctt tgagctttct caggacgtga gcttagctga
     5341 gacttctcca agaggggctg gagccctggt ctctgaggca cagtgggcct gggaaacagg
     5401 caagagtctg gttccaaagt ttttcccagc ccaggcctct gccttggcaa gcaggaagca
     5461 gggccaggtg agagccggtg agacccaggc actcgaggct agcgagctcg tttaaacaac
     5521 tagaattaca cggcgatctt gccgcccttc ttggccttaa tgagaatctc gcggatcttg
     5581 cgggcgtcca acttgccggt cagtccttta ggcacctcgt ccacgaacac aacaccaccg
     5641 cgcagcttct tggcggttgt aacctggctg gccacatagt ccacgatctc cttctcggtc
     5701 atggttttac cgtgttccag cacgacgact gcggcgggca gctcgccggc atcgtcgtcg
     5761 ggcaggccgg cgaccccggc gtcgaagatg ttggggtgtt gcagcaggat gctctccagt
     5821 tcggctgggg ctacctggta gcccttgtat ttgatcaggc tcttcagccg gtccacgatg
     5881 aagaagtgct cgtcctcgtc ccagtaggcg atgtcgccgc tgtgcagcca gccgtccttg
     5941 tcgatgagag cgtttgtagc ctcggggttg ttaacgtagc cgctcatgat catggggcca
     6001 cggacgcaca gctcgccgcg ctggttcaca cccagtgtct taccggtgtc caagtccacc
     6061 accttagcct cgaagaaggg caccaccttg cctactgcgc caggcttgtc gtccccttcg
     6121 ggggtgatca gaatggcgct ggttgtttct gtcaggccgt agccctggcg gatgcctggt
     6181 aggtggaagc gtttggccac ggcctcacct acctccttgc tgagcggcgc cccgccgctg
     6241 gcgatctcgt gcaagttgct taggtcgtac ttgtcgatga gagtgctctt agcgaagaag
     6301 ctaaatagtg tgggcaccag cagggcagat tgaatcttat agtcttgcaa gctgcgcaag
     6361 aatagctcct cctcgaagcg gtacatgagc acgacccgaa agccgcagat caagtagccc
     6421 agcgtggtga acatgccgaa gccgtggtga aatggcacca cgctgaggat agcggtgtcg
     6481 gggatgatct ggttgccgaa gatggggtcg cgggcatgac tgaatcggac acaagcggtg
     6541 cggtgcggta gggctacgcc cttgggcaat ccggtactgc cactactgtt catgatcagg
     6601 gcgatggttt tgtcccggtc gaagctctcg ggcacgaagt cgtactcgtt gaagccgggt
     6661 ggcaaatggg aagtcacgaa ggtgtacatg ctttggaagc cctggtagtc ggtcttgcta
     6721 tccatgatga tgatcttttg tatgatcggt agcttctttt gcacgttgag gatcttttgc
     6781 agccctttct tgctcacgaa tacgacggtg ggctggctga tgcccatgct gttcagcagc
     6841 tcgcgctcgt tgtagatgtc gttagctggg gccacagcca caccgatgaa cagggcaccc
     6901 aacacgggca tgaagaactg caagctattc tcgctgcaca ccacgatccg atggtttgta
     6961 ttcagcccat agcgcttcat agcttctgcc agccgaacgc tcatctcgaa gtactcggcg
     7021 taggtaatgt ccacctcgat atgtgcgtcg gtaaaggcga tggtgccggg caccagggcg
     7081 tagcgcttca tggctttgtg cagctgctcg ccggcggtcc cgtcttcgag tgggtagaat
     7141 ggcgctgggc ccttcttaat gtttttggca tcttccatgg tggctttacc aacagtaccg
     7201 gattgccaag cttgggctgc aggtcgaaag gcccggagat gaggaagagg agaacagcgc
     7261 ggcagacgtg cgcttttgaa gcgtgcagaa tgccgggcct ccggaggacc ttcgggcgcc
     7321 cgccccgccc ctgagcccgc ccctgagccc gcccccggac ccaccccttc ccagcctctg
     7381 agcccagaaa gcgaaggagc aaagctgcta ttggccgctg ccccaaaggc ctacccgctt
     7441 ccattgctca gcggtgctgt ccatctgcac gagactagtg agacgtgcta cttccatttg
     7501 tcacgtcctg cacgacgcga gctgcggggc gggggggaac ttcctgacta ggggaggagt
     7561 agaaggtggc gcgaaggggc caccaaagaa cggagccggt tggcgcctac cggtggatgt
     7621 ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa
     7681 agcgcatgct ccagactgcc ttgggaaaag cgcctcccct acccggtaga tctcgatcct
     7741 ctacgccgga cgcatcgtgg ccggagtact gcggtttatc tcgatggcta cgagggcaga
     7801 cagtaagtgg atttaccata atcccttaat tgtacgcacc gctaaaacgc gttcagcgcg
     7861 atcacggcag cagacaggta aaaatggcaa caaaccaccc gaaaaactgc cgcaatcgcg
     7921 cctgataaat tttaaccgta tgaataccta tgcaaccaga gggtacaggc cacattaccc
     7981 ccacttaatc cactgaagct gccatttttc atggtttcac catcccagcg aagggccatc
     8041 cagcgtgcgt tcctgtattt ccggactata aaaggatcta ggtgaagatc ctttttgata
     8101 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8161 aaaagatcaa aggatcttc
//
