LOCUS       sequence-469264-        8960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the 3' UTR in regulating TCTA-P2 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  TITLE     Studying the role and mechanism of the 3' UTR in regulating gene 
            function. Batch 3.
REFERENCE   2  (bases 1 to 8960)
  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: TCTA-P2
FEATURES             Location/Qualifiers
     source          1..8960
                     /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(6307..7959)
                     /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      7956..7965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8006..8505)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8054..8073)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     8447..8470
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    8562..8845
                     /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 gtcgacttgc acttccgtgt ttaataagcc
     4681 acatcctcag ttgagcctgg ggtgaaatgt gagatcctga ctctgtgcag tagtattagt
     4741 gggtgggcca ggggctgtga ataacatcat cctcagtaca gctgcaattc cagggcccct
     4801 ctaccacaaa gatggcttaa gcaaaggcag ccagatggaa gtatgatatc caacaggaag
     4861 gaagtaggca ggggtcacta aagtggctgg tggcccagca gatggaaaca gaagtatggc
     4921 cccgagggaa ggaggcaggt ccagggctac agtgctttca ggtactggtg tttctatagg
     4981 ggcatttgcc acccacatct ttggaaactc ccctggccta ttgtgacatg gcagggctgc
     5041 ctggttcttg aaggtagaga aaatgctagt ggggaggagc tgagcctgtg acgtgcattt
     5101 gtaccaggtc cagatgggaa atgatgaggc tgcaggctat gatggggcca ctgtgcacta
     5161 aatgcttggt accatcctat gaggggacaa ggtgcagacc cacattccca taggcactta
     5221 agcagtggtg gataggaggc taccccaggg gttcagctgt ccttcaggta atcagcttac
     5281 ccccaagaag gactgagagg aatatgagta ggacagttgg cagatgacaa tttcctttgc
     5341 ctggctcacg ggagaagtct ggttgggcca ggtatgtggg tgtttggtgc ctccatctgg
     5401 atagtgccag caggtacacc accctgcctt atacctgaat ggccagctaa ataaccatag
     5461 agcttaaggt ctggctcagg tcaggattcc ctggctcctt gccaacaaag caaaaccaga
     5521 gattcagaaa actcagggcc taggagaaaa cagacctgcc ttacaaaccc cagagctggt
     5581 caacctcagg cttcctctca ggaaaaaaat tttttttttg agatgggtct tgctctgttg
     5641 cccaggctgg agtacaacgg cgcaatcttg gctcactgca acctctgcct cccaggttca
     5701 agcaaatctc ctgcttcagc ctcctgagta gctgggatta ctggtgtgtg ccaccacacc
     5761 cggctaattt ttgtattttt agtagagatg gggttttcac catgttggtc aggctggtct
     5821 cgaactcctg acctcgtgat ctgcctgcct cagcctccca aagtgttggg attacaggcg
     5881 tgagccacca tgcctggcca actcaggaaa aatttttatg gggtcacatc ttggggatgg
     5941 aagagggaca ggaaaagaaa cctaagacaa aagaactgat ggtcttccca acccagaggc
     6001 ttcctgagtt atcacccctg agagaggaag aacacaatct ctctccttaa ggcacccaga
     6061 taagagtaca ggcaaactct tcacttgccc agtgcctggg tgagggagca ggcttaagag
     6121 gcagaagagg cccagccctg taggcactgg gtactccctg ctaccaaccc tgaacccccg
     6181 gcagttgtcc ctgaggtctg gggagtagca gaacccagtg tagaagccag cagacccagt
     6241 gatgcccttg gagaccctct gctgccttcc cactcgaggc tagcgagctc gtttaaacaa
     6301 ctagaattac acggcgatct tgccgccctt cttggcctta atgagaatct cgcggatctt
     6361 gcgggcgtcc aacttgccgg tcagtccttt aggcacctcg tccacgaaca caacaccacc
     6421 gcgcagcttc ttggcggttg taacctggct ggccacatag tccacgatct ccttctcggt
     6481 catggtttta ccgtgttcca gcacgacgac tgcggcgggc agctcgccgg catcgtcgtc
     6541 gggcaggccg gcgaccccgg cgtcgaagat gttggggtgt tgcagcagga tgctctccag
     6601 ttcggctggg gctacctggt agcccttgta tttgatcagg ctcttcagcc ggtccacgat
     6661 gaagaagtgc tcgtcctcgt cccagtaggc gatgtcgccg ctgtgcagcc agccgtcctt
     6721 gtcgatgaga gcgtttgtag cctcggggtt gttaacgtag ccgctcatga tcatggggcc
     6781 acggacgcac agctcgccgc gctggttcac acccagtgtc ttaccggtgt ccaagtccac
     6841 caccttagcc tcgaagaagg gcaccacctt gcctactgcg ccaggcttgt cgtccccttc
     6901 gggggtgatc agaatggcgc tggttgtttc tgtcaggccg tagccctggc ggatgcctgg
     6961 taggtggaag cgtttggcca cggcctcacc tacctccttg ctgagcggcg ccccgccgct
     7021 ggcgatctcg tgcaagttgc ttaggtcgta cttgtcgatg agagtgctct tagcgaagaa
     7081 gctaaatagt gtgggcacca gcagggcaga ttgaatctta tagtcttgca agctgcgcaa
     7141 gaatagctcc tcctcgaagc ggtacatgag cacgacccga aagccgcaga tcaagtagcc
     7201 cagcgtggtg aacatgccga agccgtggtg aaatggcacc acgctgagga tagcggtgtc
     7261 ggggatgatc tggttgccga agatggggtc gcgggcatga ctgaatcgga cacaagcggt
     7321 gcggtgcggt agggctacgc ccttgggcaa tccggtactg ccactactgt tcatgatcag
     7381 ggcgatggtt ttgtcccggt cgaagctctc gggcacgaag tcgtactcgt tgaagccggg
     7441 tggcaaatgg gaagtcacga aggtgtacat gctttggaag ccctggtagt cggtcttgct
     7501 atccatgatg atgatctttt gtatgatcgg tagcttcttt tgcacgttga ggatcttttg
     7561 cagccctttc ttgctcacga atacgacggt gggctggctg atgcccatgc tgttcagcag
     7621 ctcgcgctcg ttgtagatgt cgttagctgg ggccacagcc acaccgatga acagggcacc
     7681 caacacgggc atgaagaact gcaagctatt ctcgctgcac accacgatcc gatggtttgt
     7741 attcagccca tagcgcttca tagcttctgc cagccgaacg ctcatctcga agtactcggc
     7801 gtaggtaatg tccacctcga tatgtgcgtc ggtaaaggcg atggtgccgg gcaccagggc
     7861 gtagcgcttc atggctttgt gcagctgctc gccggcggtc ccgtcttcga gtgggtagaa
     7921 tggcgctggg cccttcttaa tgtttttggc atcttccatg gtggctttac caacagtacc
     7981 ggattgccaa gcttgggctg caggtcgaaa ggcccggaga tgaggaagag gagaacagcg
     8041 cggcagacgt gcgcttttga agcgtgcaga atgccgggcc tccggaggac cttcgggcgc
     8101 ccgccccgcc cctgagcccg cccctgagcc cgcccccgga cccacccctt cccagcctct
     8161 gagcccagaa agcgaaggag caaagctgct attggccgct gccccaaagg cctacccgct
     8221 tccattgctc agcggtgctg tccatctgca cgagactagt gagacgtgct acttccattt
     8281 gtcacgtcct gcacgacgcg agctgcgggg cgggggggaa cttcctgact aggggaggag
     8341 tagaaggtgg cgcgaagggg ccaccaaaga acggagccgg ttggcgccta ccggtggatg
     8401 tggaatgtgt gcgaggccag aggccacttg tgtagcgcca agtgcccagc ggggctgcta
     8461 aagcgcatgc tccagactgc cttgggaaaa gcgcctcccc tacccggtag atctcgatcc
     8521 tctacgccgg acgcatcgtg gccggagtac tgcggtttat ctcgatggct acgagggcag
     8581 acagtaagtg gatttaccat aatcccttaa ttgtacgcac cgctaaaacg cgttcagcgc
     8641 gatcacggca gcagacaggt aaaaatggca acaaaccacc cgaaaaactg ccgcaatcgc
     8701 gcctgataaa ttttaaccgt atgaatacct atgcaaccag agggtacagg ccacattacc
     8761 cccacttaat ccactgaagc tgccattttt catggtttca ccatcccagc gaagggccat
     8821 ccagcgtgcg ttcctgtatt tccggactat aaaaggatct aggtgaagat cctttttgat
     8881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8941 gaaaagatca aaggatcttc
//
