LOCUS       sequence-110896-        4170 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 4170)
  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 4170)
  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-FS
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(1878..1902)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1929..1945)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1929..1945)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1982..2002)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1982..2000)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2021..2037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2021..2037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2034..2056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2045..2061
                     /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(2069..2075,2076..2093,2094..2099))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2114..2135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2252..2269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2423..3011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2503..2522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3182..3973,3974..4042))
                     /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     3805..3824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4043..4147)
                     /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 tgcaaggttc aagcctactc
      781 aactgctgga gggaaggtgt ggctgtcagt acttttcatt ttccgaatcc tgctgctggg
      841 gacagcggtt gagtcagcct ggggagatga gcagtctgcc tttcgttgta acactcagca
      901 acctggttgt gaaaatgtct gctatgacaa gtctttccca atctctcatg tgcgcttctg
      961 ggtcctgcag atcatatttg tgtctgtacc cacactcttg tacctggctc atgtgttcta
     1021 tgtgatgcga aaggaagaga aactgaacaa gaaagaggaa gaactcaagg ttgcccaaac
     1081 tgatggtgtc aatgtggaca tgcacttgaa gcagattgag ataaagaagt tcaagtacgg
     1141 tattgaagag catggtaagg tgaaaatgcg aggggggttg ctgcgaacct acatcatcag
     1201 tatcctcttc aagtctatct ttgaggtggc cttcttgctg atccagtggt acatctatgg
     1261 attcagcttg agtgctgttt acacttgcaa aagagatccc tgcccacatc aggtggactg
     1321 tttcctctct cgccccacgg agaaaaccat cttcatcatc ttcatgctgg tggtgtcctt
     1381 ggtgtccctg gccttgaata tcattgaact cttctatgtt ttcttcaagg gcgttaagga
     1441 tcgggttaag ggaaagagcg acccttacca tgcgaccagt ggtgcgctga gccctgccaa
     1501 agactgtggg tctcaaaaat atgcttattt caatggctgc tcctcaccaa ccgctcccct
     1561 ctcgcctatg tctcctcctg ggtacaagct ggttactggc gacagaaaca attcttcttg
     1621 ccgcaattac aacaagcaag caagtgagca aaactgggct aattacagtg cagaacaaaa
     1681 tcgaatgggg caggcgggaa gcaccatctc taactcccat gcacagcctt ttgatttccc
     1741 cgatgataac cagaattcta aaaaactagc tgctggacat gaattacagc cactagccat
     1801 tgtggaccag cgaccttcaa gcagagccag cagtcgtgcc agcagcagac ctcggcctga
     1861 tgacctggag atctagtacc cagctttctt gtacaaagtg gttgatatcg aattcctgca
     1921 gcccggggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     1981 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2041 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2101 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2161 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2221 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2281 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2341 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2401 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2461 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2521 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2581 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2641 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2701 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2761 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2821 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2881 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2941 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3001 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3061 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3121 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3181 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3241 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3301 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3361 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3421 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3481 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3541 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3601 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3661 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3721 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3781 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3841 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3901 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3961 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4021 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4081 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4141 ttccgcgcac atttccccga aaagtgccac
//
