LOCUS       sequence-8541-ex        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  TITLE     Other Plasmids
REFERENCE   2  (bases 1 to 6056)
  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: M70 pGL3 Sp5l P1203
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1320..1329
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1326..2978
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1392..1410)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3019..3140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3030..3049)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3084..3103
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3388..3405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3559..4147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3639..3658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4318..5109,5110..5178))
                     /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     4941..4960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5179..5283)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5310..5765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5397..5416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5607..5628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5896..5944
                     /note="synthetic polyadenylation signal"
     misc_feature    5958..6049
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5998..6017
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagacg ttaagcacgt ttcctacgtg
       61 tcattttaat tctccatcac tgtggtatgt tttagcagac taaagcgatg catctgctgg
      121 gacaaaggtc gattgttatg cgactagagg tgcgaggcaa cacaaagcag tgaggatttt
      181 ctatgtatta agatgttaaa tggctttccg acttcaatgt aaattctcta agagaaaggc
      241 tcaaagtgtt agatgaactt tggatgctct gcatttaatc cctttaaacg tgatgacgcc
      301 cttaacttga acccggagtg tgaaaatatc acgcagggct ctttatttat aaaatcactt
      361 aagagtgcga actactgaat gtgcatggag gtacatatcg cgtttattaa tgagaaaaga
      421 agaatagctt aattcgatcg ttttcaaatg tcgtttagcg tcatcattaa tatgacatct
      481 tttagaacga ccagcttaca actaataaca gctaaagagg caggttcaca gatgctcata
      541 tggatcaaat acaggcctag tgaaagcaaa atccataacg cactttacaa aatagcttag
      601 tatcacttta aacccaacat gaggcctaac aataccaaca atgtatttgc ctatagtgaa
      661 tcttgaatgc cacattcatg ttacagacct attttcatta tcatttgaag tgcacgactg
      721 catcaaacta caataaaata aactaatctt aaatttaaaa gtaaagattt tggagattca
      781 ttgcacctaa tctattgaaa agtatttatt ttagtgtaaa agaaagtgtg gcttaaataa
      841 aacggtgtta gtttgaataa aaatgttata aatactgatt ataacaagct cgtccgtttg
      901 tttagttttt aatttgtcac taaagaacga gtcgttaaac ttgtaaatat cagatacatg
      961 taaacaaact ctgtcatatt aagactcttc tcacctagaa gcgctaaagg gaaggggttc
     1021 tcacaggtta acatttggca aaattaaatt agaatgaaac gaaggttata atacacaccc
     1081 ctcttcctaa tctcccaaag attgccctcc ccaaatattc ccccaccgta tctctcctcc
     1141 attcactggc cctcctgata catacttaag agaccccgtc gagaggatct tttccctata
     1201 aagagaagat gcaatccgac cgcctttgaa atgatcctca ggacttattg cgacaggccc
     1261 ttcagtcgca cttcaaaagg aagagacctg aagcttggca ttccggtact gttggtaaag
     1321 ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag
     1381 atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa
     1441 caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa
     1501 tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg
     1561 tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg
     1621 gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg
     1681 gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg
     1741 tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc
     1801 agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat
     1861 acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct
     1921 ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat
     1981 tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa
     2041 gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg
     2101 gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg
     2161 attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca
     2221 ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc
     2281 tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag
     2341 gatatgggct cactgagact acatcagcta ttctgattac acccgagggg gatgataaac
     2401 cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg
     2461 ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt
     2521 ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac
     2581 attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga
     2641 agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc
     2701 tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg
     2761 aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg
     2821 tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg
     2881 tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc
     2941 tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc
     3001 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3061 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3121 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3181 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     3241 aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     3301 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     3361 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5281 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5761 aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5821 ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt
     5881 acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     5941 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     6001 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
