LOCUS       Exported                6099 bp ds-DNA     circular SYN 31-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  AUTHORS   Westmeyer Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 31, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6099
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             964..1167
                     /codon_start=1
                     /label=MmMT3
                     /translation="MDPETCPCPTGGSCTCSDKCKCKGCKCTNCKKSCCSCCPAGCEKC
                     AKDCVCKGEEGAKAEAEKCSCCQ"
     CDS             1168..1182
                     /codon_start=1
                     /label=Linker
                     /translation="GSGPG"
     CDS             1183..2025
                     /codon_start=1
                     /label=EncBt
                     /translation="NKSQLYPDSPLTDQDFNQLDQTVIEAARRQLVGRRFIELYGPLGR
                     GMQSVFNDIFMESHEAKMDFQGSFDTEVESSRRVNYTIPMLYKDFVLYWRDLEQSKALD
                     IPIDFSVAANAARDVAFLEDQMIFHGSKEFDIPGLMNVKGRLTHLIGNWYESGNAFQDI
                     VEARNKLLEMNHNGPYALVLSPELYSLLHRVHKDTNVLEIEHVRELITAGVFQSPVLKG
                     KSGVIVNTGRNNLDLAISEDFETAYLGEEGMNHPFRVYETVVLRIKRPAAICTLIDPEE
                     "
     CDS             2026..2034
                     /codon_start=1
                     /label=Linker
                     /translation="GSG"
     CDS             2035..2058
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2112..2336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2382..2810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2824..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3202..3249
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3268..3642
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3772..3893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3966..3982
                     /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(3990..4020)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4035..4056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4344..4932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5103..5963)
                     /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(5964..6068)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggacc ctgagacatg cccttgtcca acaggcggca gctgcacctg tagcgacaag
     1021 tgcaagtgca aaggctgcaa gtgtaccaac tgcaagaagt cctgctgcag ctgctgtcct
     1081 gccggctgtg aaaagtgcgc caaggactgc gtgtgcaagg gcgaagaagg cgccaaagcc
     1141 gaggccgaga agtgctcttg ttgtcagggc agcggcccgg gcaacaagag ccagctgtac
     1201 cctgacagcc ctctgaccga ccaggacttc aaccagctgg accagaccgt gattgaagcc
     1261 gccagaaggc agctcgtcgg cagaagattc atcgagctgt acggccctct cggcagaggc
     1321 atgcagagcg tgttcaacga catcttcatg gaaagccacg aggccaagat ggacttccag
     1381 ggcagcttcg acaccgaggt ggaaagcagc agaagagtga actacacaat ccccatgctg
     1441 tacaaggact tcgtgctgta ctggcgcgac ctggaacaga gcaaggccct ggacatcccc
     1501 atcgattttt ccgtggccgc caacgccgct agagatgtgg cctttctgga agatcagatg
     1561 atctttcacg gcagcaaaga gttcgacatc cccggcctga tgaacgtgaa gggcagactg
     1621 acacacctga tcggcaattg gtacgagagc ggcaacgcct tccaggacat cgtggaagcc
     1681 cggaacaagc tgctggaaat gaaccacaac ggcccctacg ctctggtgct gagccctgag
     1741 ctgtatagcc tgctgcacag agtgcacaag gacaccaacg tcctggaaat cgagcacgtg
     1801 cgcgagctga ttaccgccgg tgtctttcag agccctgtgc tgaagggaaa gtccggcgtg
     1861 atcgtgaaca ccggcagaaa caacctggac ctggccatca gcgaggactt cgagacagcc
     1921 tatctgggcg aagagggcat gaatcacccc ttccgggtgt acgagacagt ggtgctgcgg
     1981 attaagaggc ctgccgccat ctgcaccctg atcgatcctg aagaaggctc cggcgactac
     2041 aaggacgacg acgacaaatg agcggccgct cgagtctaga gggcccgttt aaacccgctg
     2101 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2161 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2221 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2281 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2341 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2401 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2461 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2521 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     2581 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2641 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2701 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     2761 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2821 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3181 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     3241 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     3301 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     3361 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     3421 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     3481 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     3541 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     3601 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     3661 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3721 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     3781 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3841 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3901 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3961 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4021 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4081 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4141 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4501 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4741 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5101 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5161 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5221 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5281 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5341 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5401 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5461 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5521 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5581 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5641 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5701 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5761 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5821 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5881 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5941 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6001 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6061 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
