psiCHECK-2-h2.83322

LOCUS       psiCHECK-2-h2.83322               6612 bp DNA     circular SYN 03-FEB-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6612)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    1681..2011
                     /label=-7
     polyA_signal    2027..2075
                     /note="synthetic polyadenylation signal"
     promoter        2084..2835
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2865..2874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2871..4523
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3830..3838
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4567..4688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4821..4925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4926..5786
                     /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"
     rep_origin      5957..6545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagcc cgggaattcg tttaaaccta gagcggccgc
     1681 aaaagataga atttaaaaag gttgagtctc tacatggatg agaaaaaaaa agccaatata
     1741 agtcttttat ataaaccaat ctcttaaaac attttaaaaa atattttatt ttaatctatt
     1801 aaaatccctt catagcccac taacattcta aaataccagc tataaacttc aagttcattt
     1861 tttagaactt tgaaagaaaa atagatatat tgcataagaa aggtgatgaa cttaataata
     1921 gttatggaaa aagatacaaa tataacaatg aaaggtcctg cagattaatt taaccattta
     1981 attaagaaaa agagagcact attacttagg tgcggccgct ggccgcaata aaatatcttt
     2041 attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt cttcggacct
     2101 cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg gggaaggaac
     2161 gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg gcacgcagaa
     2221 gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg tttggtcggc
     2281 gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt gtgacctttt
     2341 cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc gctgggtcct
     2401 aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga ctgcggtgat
     2461 attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga gcgtccgttg
     2521 ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag gcgcgaggga
     2581 ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt gctcgcgttt
     2641 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
     2701 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
     2761 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
     2821 cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc atggccgatg
     2881 ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc accgctggcg
     2941 agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt gccttcaccg
     3001 atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct gtgcgcctgg
     3061 ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg tgctctgaga
     3121 actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg gccgtggccc
     3181 ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt tctcagccta
     3241 ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag aagaagctgc
     3301 ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc ttccagagca
     3361 tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac ttcgtgccag
     3421 agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg tctaccggcc
     3481 tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct cacgcccgcg
     3541 accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg gtgccattcc
     3601 accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt cgggtggtgc
     3661 tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac aaaattcagt
     3721 ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg atcgacaagt
     3781 acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct aaggaggtgg
     3841 gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac ggcctgaccg
     3901 agacaaccag cgccattctg attaccccag agggcgacga caagcctggc gccgtgggca
     3961 aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag accctgggag
     4021 tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc tacgtgaata
     4081 accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct ggcgacattg
     4141 cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct ctgatcaagt
     4201 acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag caccctaaca
     4261 ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg cctgccgccg
     4321 tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac tatgtggcca
     4381 gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac gaggtgccca
     4441 agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc aaggctaaga
     4501 aaggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg cttcgagcag
     4561 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4621 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4681 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4741 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg
     4801 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4861 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4921 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4981 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5041 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5101 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5161 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5221 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5281 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5341 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5401 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5461 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5521 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5581 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5641 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5701 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5761 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5821 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6601 acatggctcg ac
//