LOCUS       Exported                8250 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 8250)
  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..8250
                     /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"
     primer_bind     939..963
                     /label=EcoRI_Kozak_ol_for
     primer_bind     complement(941..963)
                     /label=pcDNA_Zeo_lin_rev
     CDS             964..1677
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1696..1941
                     /codon_start=1
                     /label=G1
                     /translation="APKTITELEKKVEEIPFKKERKFNPDLAPGTEKVTREGQKGEKTI
                     TTPTLKNPLTGVIISKGEPKEEITKDPINELTEYGPE"
     CDS             1942..2085
                     /codon_start=1
                     /label=E1domian
                     /translation="TIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVT
                     KYG"
     CDS             2086..2316
                     /codon_start=1
                     /label=G2 w/o T
                     /translation="PVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITT
                     PTLKNPLTGEIISKGESKEEITKDPINELTEY"
     CDS             2317..2325
                     /codon_start=3
                     /label=G1
                     /translation="GPE"
     CDS             2326..2469
                     /codon_start=1
                     /label=E1domian
                     /translation="TIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVT
                     KYG"
     CDS             2470..2701
                     /codon_start=1
                     /label=G2 w/o T
                     /translation="PVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITT
                     PTLKNPLTGEIISKGESKEEITKDPINELTEY"
     CDS             2702..2709
                     /codon_start=1
                     /label=G1
                     /translation="PE"
     CDS             2710..2853
                     /codon_start=1
                     /label=E1domian
                     /translation="TIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVT
                     KYG"
     CDS             2854..3085
                     /codon_start=1
                     /label=G2 w/o T
                     /translation="PVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITT
                     PTLKNPLTGEIISKGESKEEITKDPINELTEY"
     CDS             3086..3093
                     /codon_start=1
                     /label=G1
                     /translation="PE"
     CDS             3094..3237
                     /codon_start=1
                     /label=E1domian
                     /translation="TIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVT
                     KYG"
     CDS             3238..3476
                     /codon_start=1
                     /label=G2 w/o T
                     /translation="PVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITT
                     PTLKNPLTGEIISKGESKEEITKDPINELTEYGP"
     CDS             3502..4206
                     /codon_start=1
                     /label=mCherry
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     4213..4233
                     /label=pcDNA_Zeo_lin_for_FS2020
     polyA_signal    4263..4487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4533..4961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4975..5305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5156..5291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5353..5400
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5419..5793
                     /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    5923..6044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6093..6109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6117..6133
                     /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(6141..6171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6186..6207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6495..7083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7254..8114)
                     /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(8115..8219)
                     /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 accatgagca agggcgaaga actgtttacc ggcgtggtgc ccatcctggt ggaactggat
     1021 ggggatgtga acggccacaa gttcagcgtc agaggcgaag gcgaagggga tgccacaaac
     1081 ggcaagctga ccctgaagtt catctgcacc accggaaagc tgcccgtgcc ttggcctaca
     1141 ctggtcacca cactgacata cggcgtgcag tgcttcagca gataccccga ccatatgaag
     1201 cggcacgact tcttcaagag cgccatgcct gagggctacg tgcaagagag aaccatcagc
     1261 ttcaaggacg acggcaccta caagaccaga gccgaagtga agttcgaggg cgacaccctg
     1321 gtcaaccgga tcgagctgaa gggcatcgat ttcaaagagg acggcaacat cctgggccac
     1381 aagcttgagt acaacttcaa cagccacaac gtgtacatca ccgccgacaa gcagaagaac
     1441 ggcatcaagg ccaacttcaa gatccggcac aacgtggaag atggcagcgt gcagctggcc
     1501 gatcactacc agcagaacac acccatcgga gatggccctg tgctgctgcc cgataaccac
     1561 tacctgagca cacagagcgt gctgagcaag gaccccaacg agaagaggga ccacatggtg
     1621 ctgctggagt ttgtgacagc cgccggaatc acccacggca tggacgaact ttacaaaggc
     1681 ggcaccggtg gcagcgcccc taagacaatt accgagctgg aaaagaaggt ggaagagatc
     1741 cccttcaaga aagagcggaa gttcaacccc gatctggccc ctggcaccga gaaagtgaca
     1801 agagaaggcc agaagggcga gaaaaccatc accacaccta cactgaagaa ccctctgacc
     1861 ggcgtgatca tctccaaggg cgagcccaaa gaggaaatca caaaggaccc tatcaacgag
     1921 ctgaccgagt acggccccga gacaattgct cctggccaca gagatgagtt cgaccccaaa
     1981 ctgcccaccg gcgagaaaga agaagtgcct ggcaagcctg gcatcaagaa ccctgagaca
     2041 ggcgacgtcg tcagacctcc tgtggacagc gtgaccaaat acggccctgt gaagggcgac
     2101 agcatcgtgg aaaaagaaga gattccgttt gagaaagaga gaaagtttaa tcccgacctc
     2161 gctccaggga cagaaaaagt cactcgcgag ggccagaaag gggaaaagac aatcacaacc
     2221 ccaacgctga aaaatcccct gaccggggag attatcagca aaggcgagag caaagaagaa
     2281 attacgaaag atcccatcaa tgaactcaca gagtacgggc ccgagacaat tgctcctggc
     2341 cacagagatg agttcgaccc caaactgccc accggcgaga aagaagaagt gcctggcaag
     2401 cctggcatca agaaccctga gacaggcgac gtcgtcagac ctcctgtgga cagcgtgacc
     2461 aaatacggcc ctgtgaaggg cgacagcatc gtggaaaaag aagagattcc gtttgagaaa
     2521 gagagaaagt ttaatcccga cctcgctcca gggacagaaa aagtcactcg cgagggccag
     2581 aaaggggaaa agacaatcac aaccccaacg ctgaaaaatc ccctgaccgg ggagattatc
     2641 agcaaaggcg agagcaaaga agaaattacg aaagatccca tcaatgaact cacagagtac
     2701 gggcccgaga caattgctcc tggccacaga gatgagttcg accccaaact gcccaccggc
     2761 gagaaagaag aagtgcctgg caagcctggc atcaagaacc ctgagacagg cgacgtcgtc
     2821 agacctcctg tggacagcgt gaccaaatac ggccctgtga agggcgacag catcgtggaa
     2881 aaagaagaga ttccgtttga gaaagagaga aagtttaatc ccgacctcgc tccagggaca
     2941 gaaaaagtca ctcgcgaggg ccagaaaggg gaaaagacaa tcacaacccc aacgctgaaa
     3001 aatcccctga ccggggagat tatcagcaaa ggcgagagca aagaagaaat tacgaaagat
     3061 cccatcaatg aactcacaga gtacgggccc gagacaattg ctcctggcca cagagatgag
     3121 ttcgacccca aactgcccac cggcgagaaa gaagaagtgc ctggcaagcc tggcatcaag
     3181 aaccctgaga caggcgacgt cgtcagacct cctgtggaca gcgtgaccaa atacggccct
     3241 gtgaagggcg acagcatcgt ggaaaaagaa gagattccgt ttgagaaaga gagaaagttt
     3301 aatcccgacc tcgctccagg gacagaaaaa gtcactcgcg agggccagaa aggggaaaag
     3361 acaatcacaa ccccaacgct gaaaaatccc ctgaccgggg agattatcag caaaggcgag
     3421 agcaaagaag aaattacgaa agatcccatc aatgaactca cagagtacgg gcctgagacg
     3481 ggctccggag ccacaggatc tgtgtccaaa ggcgaagagg acaacatggc catcatcaaa
     3541 gaatttatgc ggttcaaggt gcacatggaa ggctccgtga atggccacga gtttgagatt
     3601 gaaggcgaag gcgagggcag accctatgag ggaacacaga ccgccaagct gaaagtgacc
     3661 aaaggcggac ctctgccttt cgcctgggac attctgagcc ctcagtttat gtacggctcc
     3721 aaggcctacg tgaagcaccc cgccgatatt cccgactacc tgaagctgag cttccccgag
     3781 ggcttcaagt gggagagagt gatgaatttc gaggacggcg gcgtggtcac cgtgacacaa
     3841 gatagttctc tgcaggacgg cgagttcatc tacaaagtga agctgcgggg caccaacttt
     3901 ccctctgacg gacccgtgat gcagaaaaag accatgggct gggaagccag cagcgagaga
     3961 atgtaccctg aagatggcgc cctgaaaggg gagatcaagc agcggctgaa actgaaggat
     4021 ggcggccact acgacgccga agtcaagacc acatacaagg ccaagaaacc agtgcagctg
     4081 ccaggcgcct acaatgtgaa catcaagctg gacatcacca gccacaatga ggactacaca
     4141 atcgtggaac agtacgagag agccgagggg agacactcta ccggcggaat ggatgagctg
     4201 tacaagtgat aagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     4261 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4321 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4381 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4441 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4501 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4681 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4861 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4981 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
     5041 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5101 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5161 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5221 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5281 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5341 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     5401 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     5461 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     5521 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     5581 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     5641 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     5701 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     5761 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     5821 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5881 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5941 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6001 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6061 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6661 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8221 acatttcccc gaaaagtgcc acctgacgtc
//
