LOCUS       sequence-322415-        4816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control vector for luciferase reporter assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4816)
  AUTHORS   Hall JM, Kainth AS, Rowton MJ, Lu E, Haddad G, Keplinger AJ, 
            Scepanovic J , Perez-Cervantes C, Chan SSK, Piccirilli JA, Moskowitz
            IP, Ruthenburg AJ
  TITLE     The cardiac lncRNA Chantico directly regulates Cxcl1 chemokine 
            transcription
  JOURNAL   bioRxiv 2021.12.23.474040
REFERENCE   2  (bases 1 to 4816)
  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"; journalName: "bioRxiv 
            2021.12.23.474040"
COMMENT     Sequence Label: pGL4-23-NegCtrl
FEATURES             Location/Qualifiers
     source          1..4816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    360..408
                     /note="synthetic polyadenylation signal"
     misc_feature    422..513
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     462..481
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    559..583
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1059..1083)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        1123..1154
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1187..2839
                     /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"
     polyA_signal    2883..3004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2894..2913)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2948..2967
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3252..3269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3423..4011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3503..3522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4211..4816,1..186,187..255))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
       61 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
      121 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
      181 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
      241 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
      301 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
      361 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
      421 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
      481 ccagtgcaag tgcaggtgcc agaacatttc tctggcctaa ctggccggta cctgagctcg
      541 ctagcctcga ggatatcaac aagtttgtac aaaaaagcag gctcctgact gatccatctc
      601 aggcaaatgt accccctgat ccaatcagct gtaactacag gtcaggaaca catagcacaa
      661 agatggctgt ggacttccca gccctgagga agagaggacc aattctcaga caggaataac
      721 tgagcaaaaa tatgtcttcc aaacaccctg tcctctcaat ccctgttgtt gtttctgcct
      781 tcattcatca tttcactatg aatggctgca aacatttcct gacaacttgc ctttcttcca
      841 gccttttctt ctcaatccac tctctaccag agtgatcttt gtacaatgaa aatcgcatca
      901 tgttactttc ctaatgaaaa cccttcaaga attctctata acctctatga caaattccag
      961 acttcttagg ttggctggaa gaagtaattt actaatttga gtcctaacgc ctaatctttt
     1021 tctttaccat aatccttcca ccctaactcc agacattcac ccagctttct tgtacaaagt
     1081 ggttgatatc aagatctggc ctcggcggcc aagcttagac actagagggt atataatgga
     1141 agctcgactt ccagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
     1201 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
     1261 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
     1321 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
     1381 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
     1441 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
     1501 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
     1561 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
     1621 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
     1681 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
     1741 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
     1801 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
     1861 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
     1921 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
     1981 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
     2041 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     2101 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     2161 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     2221 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     2281 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     2341 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     2401 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     2461 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     2521 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     2581 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     2641 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     2701 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     2761 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     2821 cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat
     2881 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2941 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3001 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3061 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     3121 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3181 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3241 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3421 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3601 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3841 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc
     4201 actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc
     4261 gttcgtccat agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac
     4321 catcaggccc cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt
     4381 cagcaatgaa ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg
     4441 cctccatcca gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta
     4501 gtttccgaag agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta
     4561 tggcttcgtt caactctggt tcccagcggt caagccgggt cacatgatca cccatattat
     4621 gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg
     4681 tgttgtcgct catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa
     4741 gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc
     4801 gaccaagctg ctcttg
//
