LOCUS       sequence-110893-        4171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning, T7 RNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4171)
  AUTHORS   Salat-Canela C, Sese M, Peula C, Ramon y Cajal S, Aasen T
  TITLE     Internal translation of the connexin 43 transcript.
  JOURNAL   Cell Commun Signal. 2014 May 8;12:31. doi: 10.1186/1478-811X-12-31.
  PUBMED    24884945
REFERENCE   2  (bases 1 to 4171)
  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: "Cell Commun
            Signal."; date: "2014-05-8"; volume: "12:31. doi"; pages: " 
            10.1186/1478-811X-12-31"
COMMENT     Sequence Label: pBS-Cx43
FEATURES             Location/Qualifiers
     source          1..4171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    702..726
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1879..1903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1930..1946)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1983..2003)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1983..2001)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2022..2038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2022..2038)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2035..2057)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2046..2062
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2070..2076,2077..2094,2095..2100))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2115..2136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2253..2270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2424..3012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2504..2523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3183..3974,3975..4043))
                     /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     3806..3825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4044..4148)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc aacaagtttg tacaaaaaag
      721 caggcttcat gggtgactgg agcgccttag gcaaactcct tgacaaggtt caagcctact
      781 caactgctgg agggaaggtg tggctgtcag tacttttcat tttccgaatc ctgctgctgg
      841 ggacagcggt tgagtcagcc tggggagatg agcagtctgc ctttcgttgt aacactcagc
      901 aacctggttg tgaaaatgtc tgctatgaca agtctttccc aatctctcat gtgcgcttct
      961 gggtcctgca gatcatattt gtgtctgtac ccacactctt gtacctggct catgtgttct
     1021 atgtgatgcg aaaggaagag aaactgaaca agaaagagga agaactcaag gttgcccaaa
     1081 ctgatggtgt caatgtggac atgcacttga agcagattga gataaagaag ttcaagtacg
     1141 gtattgaaga gcatggtaag gtgaaaatgc gaggggggtt gctgcgaacc tacatcatca
     1201 gtatcctctt caagtctatc tttgaggtgg ccttcttgct gatccagtgg tacatctatg
     1261 gattcagctt gagtgctgtt tacacttgca aaagagatcc ctgcccacat caggtggact
     1321 gtttcctctc tcgccccacg gagaaaacca tcttcatcat cttcatgctg gtggtgtcct
     1381 tggtgtccct ggccttgaat atcattgaac tcttctatgt tttcttcaag ggcgttaagg
     1441 atcgggttaa gggaaagagc gacccttacc atgcgaccag tggtgcgctg agccctgcca
     1501 aagactgtgg gtctcaaaaa tatgcttatt tcaatggctg ctcctcacca accgctcccc
     1561 tctcgcctat gtctcctcct gggtacaagc tggttactgg cgacagaaac aattcttctt
     1621 gccgcaatta caacaagcaa gcaagtgagc aaaactgggc taattacagt gcagaacaaa
     1681 atcgaatggg gcaggcggga agcaccatct ctaactccca tgcacagcct tttgatttcc
     1741 ccgatgataa ccagaattct aaaaaactag ctgctggaca tgaattacag ccactagcca
     1801 ttgtggacca gcgaccttca agcagagcca gcagtcgtgc cagcagcaga cctcggcctg
     1861 atgacctgga gatctagtac ccagctttct tgtacaaagt ggttgatatc gaattcctgc
     1921 agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt
     1981 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2041 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2821 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     2881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3061 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3121 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3181 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3241 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3301 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3361 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3421 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3481 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3541 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3601 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3661 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3721 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3781 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3841 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3901 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3961 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4021 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4081 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4141 gttccgcgca catttccccg aaaagtgcca c
//
