LOCUS       pCS2plus8-CYP2A6        5720 bp DNA     circular SYN 18-SEP-2024
DEFINITION  C-terminus Empty vector  (5132 ???).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 18, 2024 from SnapGene 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     72..131
                     /label=Fwd-primer-ext
     primer_bind     115..154
                     /label=Fwd-primer-CYP2A6
     primer_bind     115..138
                     /label=Fwd-primer-CYP-33E1
     misc_feature    136..1617
                     /label=CYP2A6
     misc_feature    136..138
                     /label=START
     primer_bind     complement(1601..1642)
                     /label=Rev-primer-CYP2A6
     primer_bind     complement(1618..1674)
                     /label=Rev-Primer-EXT
     primer_bind     complement(1618..1642)
                     /label=Rev-primer-cyp-33e1
     CDS             1624..1647
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1654..1674
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1684..1701
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1769..1903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2016..2034)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2055..2071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2079..2095
                     /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(2103..2109,2110..2127,2128..2133))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2148..2169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2457..3045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3216..4007,4008..4076))
                     /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"
     promoter        complement(4077..4181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4207..4662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4736..5720
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccactagt ggcgcgccat taattaaagg ccggccagcg
      121 atcgccggac ccaccatgct ggcctcaggg atgcttctgg tggccttgct ggtctgcctg
      181 actgtgatgg tcttgatgtc tgtttggcag cagaggaaga gcaaggggaa gctgcctccg
      241 ggacccaccc cattgccctt cattggaaac tacctgcaac tgaacacaga gcagatgtac
      301 aactccctca tgaagatcag tgagcgctat ggccccgtgt tcaccattca cttggggccc
      361 cggcgggtcg tggtgctgtg tggacatgat gccgtcaggg aggctctggt ggaccaggct
      421 gaggagttca gcgggcgagg cgagcaagcc accttcgact gggtcttcaa aggctatggc
      481 gtggtattca gcaacgggga gcgcgccaag cagctccggc gcttctccat cgccaccctg
      541 cgggacttcg gggtgggcaa gcgaggcatc gaggagcgca tccaggagga ggcgggcttc
      601 ctcatcgacg ccctccgggg cactggcggc gccaatatcg atcccacctt cttcctgagc
      661 cgcacagtct ccaatgtcat cagctccatt gtctttgggg accgctttga ctataaggac
      721 aaagagttcc tgtcactgtt gcgcatgatg ctaggaatct tccagttcac gtcaacctcc
      781 acggggcagc tctatgagat gttctcttcg gtgatgaaac acctgccagg accacagcaa
      841 caggcctttc agttgctgca agggctggag gacttcatag ccaagaaggt ggagcacaac
      901 cagcgtacgc tggatcccaa ttccccacgg gacttcattg actcctttct catccgcatg
      961 caggaggagg agaagaaccc caacacggag ttctacttga aaaacctggt gatgaccacg
     1021 ttgaacctct tcattggggg caccgagacc gtcagcacca ccctgcgcta tggcttcttg
     1081 ctgctcatga agcacccaga ggtggaggcc aaggtccatg aggagattga cagagtgatc
     1141 ggcaagaacc ggcagcccaa gtttgaggac cgggccaaga tgccctacat ggaggcagtg
     1201 atccacgaga tccaaagatt tggagacgtg atccccatga gtttggcccg cagagtcaaa
     1261 aaggacacca agtttcggga tttcttcctc cctaagggca ccgaagtgta ccctatgctg
     1321 ggctctgtgc tgagagaccc cagtttcttc tccaaccccc aggacttcaa tccccagcat
     1381 ttcctgaatg agaaggggca gtttaagaag agtgatgctt ttgtgccctt ttccatcgga
     1441 aagcggaacg ctttcggaga aggcctggcc agaatggagc tctttctctt cttcaccacc
     1501 gtcatgcaga acttccgcct caagtcctcc cagtcaccta aggacattga cgtgtccccc
     1561 aaacacgtgg gctttgccac gatcccacga aactacacca tgagcttcct gccccgctca
     1621 ggggattata aagacgatga cgacaagtca ggggaaaact tgtatttcca gggcggcagc
     1681 agccatcatc atcatcatca tgggtcaggg taataatcga attcaaggcc tctcgagcct
     1741 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1801 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1861 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1921 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1981 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2041 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2101 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2161 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2221 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2281 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2341 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2401 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2461 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2521 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2581 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2641 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2701 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2761 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2821 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2881 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2941 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3001 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3061 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3121 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3181 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3241 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3301 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3361 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3421 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3481 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3541 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3601 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3661 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3721 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3781 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3841 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3901 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3961 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4021 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4081 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4141 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4201 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4261 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4321 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4381 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4441 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4501 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4561 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4621 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4681 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacca
     4741 tagccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     4801 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     4861 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acttggcact
     4921 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4981 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     5041 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     5101 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     5161 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     5221 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     5281 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     5341 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     5401 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     5461 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     5521 actattggaa ggacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     5581 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     5641 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     5701 gcaatgctcg tttagggaac
//
