LOCUS       Exported                8355 bp DNA     circular SYN 13-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8355)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 13, 2023 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8355
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    1762..5370
                     /label=S ecto dFu 2P
     misc_feature    5380..5460
                     /label=Foldon
     CDS             5467..5490
                     /label=PreScission site
     CDS             5494..5517
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5494..5511
                     /label=6His
     CDS             5497..5514
                     /label=6His
     CDS             5500..5517
                     /label=6His
     CDS             5524..5547
                     /label=Strep tag II
     CDS             5587..5610
                     /label=Strep tag II
     polyA_signal    5777..5832
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6187..6207)
                     /label=M13-rev
     primer_bind     complement(6191..6207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(6213..6235)
                     /label=LacO
     protein_bind    6215..6231
                     /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(6239..6269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(6240..6269)
                     /label=lac
     protein_bind    6284..6305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6587..7215)
                     /direction=LEFT
                     /label=ColE1 origin
     rep_origin      complement(6605..7193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7364..8224)
                     /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"
     CDS             complement(7367..8026)
                     /label=AmpR
     promoter        complement(8225..8329)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(8266..8294)
                     /label=Amp prom
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattgcg gccgtctcag
     1741 gccgagttcg gtaccgccac catgttcgtg ttcctggtgc tcctgcctct ggtgagcagc
     1801 cagtgcgtga acctgatcac ccgaacccag tcctacacca acagctttac acggggcgtg
     1861 tactaccctg acaaggtgtt cagatctagc gtcctgcaca gcactcagga cctcttcctg
     1921 ccgttcttca gcaacgtgac atggttccac gccatcagcg gcacaaacgg aaccaagcgg
     1981 tttgataacc ccgtcctgcc attcaatgat ggagtttact tcgccagtac cgagaagagt
     2041 aacatcatcc ggggctggat cttcggcacc accctggata gcaaaacaca gagcctcctg
     2101 atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg agttccagtt ttgcaatgat
     2161 cctttcctgg atgtgtacta ccacaagaac aacaagagct ggatggaaag cgagttcaga
     2221 gtctacagca gcgccaacaa ctgcacattc gagtacgtct ctcagccttt tctgatggac
     2281 cttgagggga aacaaggcaa cttcaagaac ctgagagaat tcgtgttcaa gaacatcgac
     2341 ggctacttca aaatctactc caagcacaca cccatcaacc tgggccggga cctccctcag
     2401 ggcttcagcg ccctggaacc cctggtcgac ctgcccatag gcatcaacat aacgcggttc
     2461 caaaccctgc tggccctgca tagatcctac ctgactcctg gcgacagcag cagcggatgg
     2521 accgccggag ctgcagccta ctatgtgggc tacctgcaac ctagaacctt cctgctgaag
     2581 tacaacgaga acggcacaat cacagacgcc gtcgactgcg ccctggaccc tctctctgag
     2641 acaaagtgca ccctgaagtc cttcaccgtg gaaaagggca tctaccagac cagcaacttc
     2701 cgggtgcagc ctacagagag catcgtgcga tttccaaaca ttaccaacct ctgccccttc
     2761 gacgaggtgt ttaacgccac aagatttgcc tccgtttacg cctggaatag aaagagaatc
     2821 agcaattgtg tggccgacta ctccgtgctg tataacttcg cccctttctt cgccttcaag
     2881 tgctacggcg tttccccaac aaagctgaat gacctgtgct tcaccaacgt gtacgccgac
     2941 tccttcgtaa ttagaggcaa tgaggtgtcc cagatcgcac caggccagac cggtaacatc
     3001 gctgactaca actataagct gcctgatgat tttacaggct gcgtgatcgc ctggaactcc
     3061 aacaagctgg atagcaaggt gggcggcaac tacaactaca gataccggct gtttcgcaag
     3121 tctaacctga aacctttcga gagagacatc tccacagaga tctaccaggc cggtaacaag
     3181 ccttgtaacg gggtggccgg cgtgaactgt tacttccctc tgcaaagcta cggcttccgg
     3241 cctacctatg gagtcggcca ccagccatac cgggtggtcg tgctgtcctt cgagttactc
     3301 cacgcccccg ccaccgtctg cggtcctaag aagtccacca atctggttaa gaacaaatgc
     3361 gtgaacttca acttcaacgg cctgaccggg accggcgtgc tgaccgaaag caacaaaaag
     3421 ttcctcccct tccagcagtt cggccgtgat atcgctgaca ccacagatgc cgtcagagat
     3481 ccacagaccc tggaaatcct ggatattaca ccctgctcct tcggaggagt ttctgtgatc
     3541 acccccggga ccaataccag caaccaggtg gctgtgctgt accaaggtgt taactgcacc
     3601 gaggttcctg tggccatcca cgccgatcag ctgacaccta cttggagagt gtactccact
     3661 ggctccaatg tgttccagac cagggccgga tgtctgatcg gcgccgagta cgtgaataac
     3721 agttacgagt gcgacatccc tatcggcgcc ggcatctgtg ccagctacca gacccagaca
     3781 aagagccatg ggtctgcttc ctctgtagct agccagagca tcatcgccta caccatgagc
     3841 ctgggcgcag agaacagcgt ggcctattcc aacaactcta tcgccattcc caccaacttt
     3901 acaattagcg tcacaacaga gatcctgccc gtgagcatga ccaagaccag cgtggactgt
     3961 acaatgtaca tctgtggcga cagcactgaa tgcagcaacc tgctgctgca atacggctcc
     4021 ttttgcaccc aactgaagcg ggcgctgacc ggaatcgccg tggaacagga caaaaatacc
     4081 caggaggtgt tcgcccaagt gaagcagatc tacaagaccc cacctatcaa gtacttcggc
     4141 ggctttaact ttagccagat tctccctgat ccttctaagc ctagcaagcg gagctttatc
     4201 gaggatctgc tgttcaacaa ggtcaccctg gccgatgccg gctttatcaa acagtatggc
     4261 gattgtctgg gcgacatagc cgccagagat ctgatctgcg cccagaaatt caacggcctg
     4321 acagttctcc cacctctgct gaccgacgag atgatcgctc agtacacctc tgccctgctg
     4381 gctggcacca tcacatctgg gtggacattt ggcgccggcg ccgccctgca gatccccttt
     4441 gccatgcaga tggcctatag attcaacgga atcggcgtga cccagaacgt gctgtatgaa
     4501 aaccagaagc tgatcgctaa ccagttcaat tctgccatcg gcaagatcca ggactccctc
     4561 tcctctaccg ccagcgccct gggcaaactg caggacgtgg tgaatcacaa cgcccaagcc
     4621 ctgaacaccc tggtgaagca gctcagcagc aagtttggcg ccatcagctc tgtgctgaac
     4681 gatatcctgt ctagactgga ccctccagaa gccgaagtcc agatcgatag actgatcaca
     4741 ggcagactgc agtccctgca aacctacgtg acccaacagc tgatcagggc cgctgaaata
     4801 agagccagcg ccaatctcgc cgctaccaag atgtccgagt gtgtgctggg acagtctaaa
     4861 cgcgttgact tctgcggcaa aggctatcac ctgatgagct tcccccagag cgcgccgcac
     4921 ggcgtggtgt tcctgcatgt gacatacgtg cctgcccaag agaagaattt cacaaccgcc
     4981 cctgccatct gccacgacgg caaggcccac ttcccaagag agggcgtttt cgtttccaat
     5041 ggcacacact ggttcgtgac acaaagaaac ttctacgaac cccagattat caccaccgac
     5101 aacaccttcg tgagtggcaa ttgtgacgtg gtcatcggaa tcgtgaacaa cacagtgtac
     5161 gaccctctgc aacctgagct ggactctttt aaggaagagc tggacaagta ctttaaaaac
     5221 cacaccagcc ccgatgtgga cctgggcgac atcagtggca ttaacgccag cgtggtgaac
     5281 atccaaaagg aaatcgacag actgaacgag gtggccaaga acctgaacga gtccctgatc
     5341 gacctgcagg agctcggcaa atacgagcag ggatccggat acatccccga ggcccccaga
     5401 gatggccagg cctacgtgcg gaaggacggc gagtgggtac tgctgagcac attcctgggc
     5461 agatccctgg aggtgctgtt ccagggccca ggccatcacc accatcacca ccatcatagc
     5521 gcctggtccc acccccagtt cgagaagggc ggcggtagtg gagggggcgg atctggcggc
     5581 tcagcttgga gccaccccca gttcgaaaag tgataatgac tcgagcgata attcactcct
     5641 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     5701 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     5761 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     5821 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     5881 tatttggttt agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct
     5941 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     6001 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     6061 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     6121 cccagtcata gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg
     6181 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6241 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6301 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     6361 tcgatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6781 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7021 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7141 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7201 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7261 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7321 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7441 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7561 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7801 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7861 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8041 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8221 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8281 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8341 gaaaagtgcc acctg
//
