LOCUS       Exported                6505 bp ds-DNA     circular SYN 22-FEB-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 23, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6505
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             847..2655
                     /codon_start=1
                     /label=hAT-7_PM
                     /translation="MSGSRKRKVDNECRVFNTEWTTKYFFTEVQSKAVCLICRETVAVF
                     KEYNISRHFATKHANYASKQSTQERAATAQRLAANLQTQQHFFHRQTAIQESTTKASFL
                     VAFEIAKASKPFSEGEFVKECMVQTADILCPEIKSKFEKVSLSRRTVTRRVELIDENIA
                     SQLNKKSDSFELYSLALDESTDVKDTAQLLIFIRGIDDSFAITEEFLTMESLKGTTRGE
                     DLYNQVSAVIERMKLPWSKLVNVTTDGSPNLTGKNVGLLKRIQNKVKEENPDQDLIFLH
                     CIIHQESLCKSVLQLNHVVNPAVKLVNFIRARGLQHRQFITFLEETDADHQDLLYHSRV
                     RWLSLGKVLQRVWELKEDIIAFLELMGKSDEFPELSDKNWLSDFAFAVDIFSHMNELNV
                     KLQGKDQFVHDMYKHVKAFKSKLTLFSRQIANKSFAHFPTLAMQEEAPRNAKKYSKSLE
                     DLHGEFCRRFSDFENIEQSLQLVSCPLSQDSETAPQELQLELIDLQSDSVLKEKFNSVK
                     LNDFYASLNRATFPNLRRTAQKMLTLFGSTYVCEQTFSVMNANKARHRSKLTDQHLRSI
                     LRIATTKITPDLDALAKMGDQQHCSH"
     CDS             1690..1692
                     /codon_start=1
                     /label=C265A C282A TGC-GCC
                     /translation="C"
     CDS             1699..1701
                     /codon_start=1
                     /label=H268A H285A CAC-GCC
                     /translation="H"
     CDS             1711..1713
                     /codon_start=1
                     /label=L272A L289A CTG-GCG
                     /translation="L"
     CDS             1867..1869
                     /codon_start=1
                     /label=R318A R341A CGC-GCC
                     /translation="R"
     CDS             1876..1878
                     /codon_start=1
                     /label=S321A S344A AGC-GCC
                     /translation="S"
     CDS             2296..2298
                     /codon_start=1
                     /label=P437A P484A CCG-GCG
                     /translation="P"
     CDS             2509..2511
                     /codon_start=1
                     /label=S576A S555A AGC-GCC
                     /translation="S"
     CDS             2539..2541
                     /codon_start=1
                     /label=R586A R565A CGC-GCC
                     /translation="R"
     promoter        complement(2693..2711)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2737..2961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3007..3435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3449..3778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3629..3764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3944..4065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4114..4130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4138..4154
                     /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(4162..4192)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4207..4228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5275..6135)
                     /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(6136..6240)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacgtc cgcgcagggc aatagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc
      841 gccaccatga gcggtagccg taaacgcaag gtggataacg agtgccgcgt gttcaacacc
      901 gaatggacca ccaagtattt ctttaccgag gtgcagagca aagcggtgtg tctgatctgc
      961 cgtgaaaccg tggcggtgtt caaggagtac aacattagcc gccacttcgc gactaagcat
     1021 gccaactacg cgagcaaaca gagcacccaa gaacgtgcag caaccgcgca acgtttagcg
     1081 gccaacttac agacccagca gcattttttc caccgccaga ccgcgattca agagagcacc
     1141 accaaagcca gcttcctggt ggcattcgag atagccaaag cgagcaaacc gttcagcgaa
     1201 ggcgaatttg tgaaggaatg catggtgcag accgcggata ttctgtgccc ggaaatcaaa
     1261 agcaaattcg agaaagtgag cctgagccgt cgtaccgtta cccgtcgtgt ggaactgatt
     1321 gatgagaaca ttgcgagcca gctgaacaaa aaaagcgata gctttgagct gtacagcctg
     1381 gcgctggatg aaagcaccga tgtgaaggat accgcgcagc tgctgatttt tattcgcggc
     1441 attgatgaca gctttgccat taccgaggaa tttctgacca tggaaagcct gaaaggtacc
     1501 actcgcggcg aagacctgta taaccaggtg agcgccgtga ttgaacgcat gaaactgccg
     1561 tggagcaagc tggttaacgt gaccaccgat ggtagcccta acctgaccgg caaaaacgtg
     1621 ggcctgctga aacgcattca gaacaaagtg aaagaggaga acccggatca ggatctgatt
     1681 ttcctgcatt gcatcatcca ccaggaaagc ctgtgcaaaa gcgtgctgca gctgaaccac
     1741 gttgtgaacc ctgcggtgaa actggtgaac tttattcgcg cccgcggttt acaacatcgc
     1801 cagtttatca ccttcctgga agaaaccgac gccgatcatc aggatctgct gtatcatagc
     1861 cgcgttcgct ggttaagcct gggtaaagtt ctgcagcgcg tgtgggaact gaaggaagat
     1921 atcattgcgt ttctggagct gatgggcaaa agcgatgagt ttcctgagct gagcgataaa
     1981 aactggctga gcgatttcgc attcgcggtg gacatcttca gccatatgaa cgaactgaac
     2041 gtgaaactgc agggcaaaga tcagtttgtg cacgacatgt ataagcacgt gaaggccttt
     2101 aaaagcaaac tgaccctgtt tagccgccag attgccaaca aaagcttcgc ccatttccct
     2161 accctggcga tgcaagaaga agcaccgcgc aacgcgaaaa agtatagcaa aagcctggaa
     2221 gatctgcatg gcgaattttg ccgccgcttt agcgattttg aaaacatcga acagagcctg
     2281 cagttagtga gctgtccgtt aagccaggat agcgaaaccg cgcctcagga attacagctg
     2341 gaactgattg atctgcagag cgatagcgtg ctgaaagaga aattcaacag cgtgaagctg
     2401 aacgactttt atgcgagcct gaaccgtgcg acctttccta atctgcgccg taccgcccaa
     2461 aaaatgctga ccctgttcgg cagcacctat gtgtgcgagc agaccttcag cgtgatgaac
     2521 gccaacaaag cgcgccatcg cagcaaactg accgatcagc atctgcgcag catcttacgc
     2581 attgcgacca ccaaaatcac ccctgatctg gatgcgctgg ccaaaatggg tgaccagcag
     2641 cattgcagcc attaatgagc ggccgctcga gcatgcatct agagggccct attctatagt
     2701 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     2761 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2821 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2881 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2941 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3001 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3061 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3421 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3781 gctccccgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3841 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3901 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     3961 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4021 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4081 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4141 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4201 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4261 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4321 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6241 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6301 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     6361 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     6421 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     6481 gatgtacggg ccagatatac gcgtt
//
