LOCUS       Exported                8358 bp DNA     circular SYN 14-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8358)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 20, 2023 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8358
                     /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..5373
                     /label=S ecto dFu 2P
     misc_feature    5383..5463
                     /label=Foldon
     CDS             5470..5493
                     /label=PreScission site
     CDS             5497..5520
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5497..5514
                     /label=6His
     CDS             5500..5517
                     /label=6His
     CDS             5503..5520
                     /label=6His
     CDS             5527..5550
                     /label=Strep tag II
     CDS             5590..5613
                     /label=Strep tag II
     polyA_signal    5780..5835
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6190..6210)
                     /label=M13-rev
     primer_bind     complement(6194..6210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(6216..6238)
                     /label=LacO
     protein_bind    6218..6234
                     /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(6242..6272)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(6243..6272)
                     /label=lac
     protein_bind    6287..6308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6590..7218)
                     /direction=LEFT
                     /label=ColE1 origin
     rep_origin      complement(6608..7196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7367..8227)
                     /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(7370..8029)
                     /label=AmpR
     promoter        complement(8228..8332)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(8269..8297)
                     /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 gccatccacg tgagcggcac aaacggaacc
     1981 aagcggtttg ataaccccgc cctgccattc aatgatggag tttacttcgc cagtaccgag
     2041 aagagtaaca tcatccgggg ctggatcttc ggcaccaccc tggatagcaa aacacagagc
     2101 ctcctgatcg tgaacaatgc cacgaacgtc gtgatcaagg tgtgcgagtt ccagttttgc
     2161 aatgatcctt tcctggacgt gtaccagaag aacaacaaga gctggatgga aagcgagttc
     2221 agagtctaca gcagcgccaa caactgcaca ttcgagtacg tctctcagcc ttttctgatg
     2281 gaccttgagg ggaaagaagg caacttcaag aacctgagag aattcgtgtt caagaacatc
     2341 gacggctact tcaaaatcta ctccaagcac acacccatca acctggagcg ggacctccct
     2401 cagggcttca gcgccctgga acccctggtc gacctgccca taggcatcaa cataacgcgg
     2461 ttccaaaccc tgctggccct gcatagatcc tacctgactc ctgtggacag cagcagcgga
     2521 tggaccgccg gagctgcagc ctactatgtg ggctacctgc aacctagaac cttcctgctg
     2581 aagtacaacg agaacggcac aatcacagac gccgtcgact gcgccctgga ccctctctct
     2641 gagacaaagt gcaccctgaa gtccttcacc gtggaaaagg gcatctacca gaccagcaac
     2701 ttccgggtgc agcctacaga gagcatcgtg cgatttccaa acattaccaa cctctgcccc
     2761 ttccacgagg tgtttaacgc cacaacattt gcctccgttt acgcctggaa tagaaagaga
     2821 atcagcaatt gtgtggccga ctactccgtg atctataact tcgccccttt cttcgccttc
     2881 aagtgctacg gcgtttcccc aacaaagctg aatgacctgt gcttcaccaa cgtgtacgcc
     2941 gactccttcg taattagagg caatgaggtg tcccagatcg caccaggcca gaccggtaac
     3001 atcgctgact acaactataa gctgcctgat gattttacag gctgcgtgat cgcctggaac
     3061 tccaacaagc tggatagcaa gcctagcggc aactacaact acctgtaccg gctgtttcgc
     3121 aagtctaagc tgaaaccttt cgagagagac atctccacag agatctacca ggccggtaac
     3181 aagccttgta acggggtggc cggccccaac tgttactccc ctctgcaaag ctacggcttc
     3241 cggcctacct atggagtcgg ccaccagcca taccgggtgg tcgtgctgtc cttcgagtta
     3301 ctccacgccc ccgccaccgt ctgcggtcct aagaagtcca ccaatctggt taagaacaaa
     3361 tgcgtgaact tcaacttcaa cggcctgacc gggaccggcg tgctgaccga aagcaacaaa
     3421 aagttcctcc ccttccagca gttcggccgt gatatcgctg acaccacaga tgccgtcaga
     3481 gatccacaga ccctggaaat cctggatatt acaccctgct ccttcggagg agtttctgtg
     3541 atcacccccg ggaccaatac cagcaaccag gtggctgtgc tgtaccaagg tgttaactgc
     3601 accgaggttc ctgtggccat ccacgccgat cagctgacac ctacttggag agtgtactcc
     3661 actggctcca atgtgttcca gaccagggcc ggatgtctga tcggcgccga gtacgtgaat
     3721 aacagttacg agtgcgacat ccctatcggc gccggcatct gtgccagcta ccagacccag
     3781 acaaagagcc atagacgggc tagatctgta gctagccaga gcatcatcgc ctacaccatg
     3841 agcctgggcg cagagaacag cgtggcctat tccaacaact ctatcgccat tcccaccaac
     3901 tttacaatta gcgtcacaac agagatcctg cccgtgagca tgaccaagac cagcgtggac
     3961 tgtacaatgt acatctgtgg cgacagcact gaatgcagca acctgctgct gcaatacggc
     4021 tccttttgca cccaactgaa gcgggcgctg accggaatcg ccgtggaaca ggacaaaaat
     4081 acccaggagg tgttcgccca agtgaagcag atctacaaga ccccacctat caagtacttc
     4141 ggcggcttta actttagcca gattctccct gatccttcta agcctagcaa gcggagcttt
     4201 atcgaggatc tgctgttcaa caaggtcacc ctggccgatg ccggctttat caaacagtat
     4261 ggcgattgtc tgggcgacat agccgccaga gatctgatct gcgcccagaa attcaacggc
     4321 ctgacagttc tcccacctct gctgaccgac gagatgatcg ctcagtacac ctctgccctg
     4381 ctggctggca ccatcacatc tgggtggaca tttggcgccg gcgccgccct gcagatcccc
     4441 tttgccatgc agatggccta tagattcaac ggaatcggcg tgacccagaa cgtgctgtat
     4501 gaaaaccaga agctgatcgc taaccagttc aattctgcca tcggcaagat ccaggactcc
     4561 ctctcctcta ccgccagcgc cctgggcaaa ctgcaggacg tggtgaatca caacgcccaa
     4621 gccctgaaca ccctggtgaa gcagctcagc agcaagtttg gcgccatcag ctctgtgctg
     4681 aacgatatcc tgtctagact ggacaaggtg gaagccgaag tccagatcga tagactgatc
     4741 acaggcagac tgcagtccct gcaaacctac gtgacccaac agctgatcag ggccgctgaa
     4801 ataagagcca gcgccaatct cgccgctacc aagatgtccg agtgtgtgct gggacagtct
     4861 aaacgcgttg acttctgcgg caaaggctat cacctgatga gcttccccca gagcgcgccg
     4921 cacggcgtgg tgttcctgca tgtgacatac gtgcctgccc aagagaagaa tttcacaacc
     4981 gcccctgcca tctgccacga cggcaaggcc cacttcccaa gagagggcgt tttcgtttcc
     5041 aatggcacac actggttcgt gacacaaaga aacttctacg aaccccagat tatcaccacc
     5101 gacaacacct tcgtgagtgg caattgtgac gtggtcatcg gaatcgtgaa caacacagtg
     5161 tacgaccctc tgcaacctga gctggactct tttaaggaag agctggacaa gtactttaaa
     5221 aaccacacca gccccgatgt ggacctgggc gacatcagtg gcattaacgc cagcgtggtg
     5281 aacatccaaa aggaaatcga cagactgaac gaggtggcca agaacctgaa cgagtccctg
     5341 atcgacctgc aggagctcgg caaatacgag cagggatccg gatacatccc cgaggccccc
     5401 agagatggcc aggcctacgt gcggaaggac ggcgagtggg tactgctgag cacattcctg
     5461 ggcagatccc tggaggtgct gttccagggc ccaggccatc accaccatca ccaccatcat
     5521 agcgcctggt cccaccccca gttcgagaag ggcggcggta gtggaggggg cggatctggc
     5581 ggctcagctt ggagccaccc ccagttcgaa aagtgataat gactcgagcg ataattcact
     5641 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     5701 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     5761 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     5821 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     5881 gagtatttgg tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg
     5941 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     6001 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     6061 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     6121 actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct
     6181 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6241 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6301 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6361 ggatcgatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6421 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6781 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     6841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7021 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8341 cccgaaaagt gccacctg
//
