LOCUS       Exported                6558 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 6558)
  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..6558
                     /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..1185
                     /codon_start=1
                     /label=Linker
                     /translation="GGTGGS"
     CDS             1186..1350
                     /codon_start=1
                     /label=SeSmtA
                     /translation="TSTTLVKCACEPCLCNVDPSKAIDRNGLYYCSEACADGHTGGSKG
                     CGHTGCNCHG"
     CDS             1360..1605
                     /codon_start=1
                     /label=TaEC1
                     /translation="GCDDKCGCAVPCPGGTGCRCTSARSGAAAGEHTTCGCGEHCGCNP
                     CACGREGTPSGRANRRANCSCGAACNCASCGSATAPG"
     CDS             1606..1641
                     /codon_start=1
                     /label=Linker
                     /translation="GGGSGGGSGGSA"
     CDS             1642..2484
                     /codon_start=1
                     /label=EncBt
                     /translation="NKSQLYPDSPLTDQDFNQLDQTVIEAARRQLVGRRFIELYGPLGR
                     GMQSVFNDIFMESHEAKMDFQGSFDTEVESSRRVNYTIPMLYKDFVLYWRDLEQSKALD
                     IPIDFSVAANAARDVAFLEDQMIFHGSKEFDIPGLMNVKGRLTHLIGNWYESGNAFQDI
                     VEARNKLLEMNHNGPYALVLSPELYSLLHRVHKDTNVLEIEHVRELITAGVFQSPVLKG
                     KSGVIVNTGRNNLDLAISEDFETAYLGEEGMNHPFRVYETVVLRIKRPAAICTLIDPEE
                     "
     CDS             2485..2493
                     /codon_start=1
                     /label=Linker
                     /translation="GSG"
     CDS             2494..2517
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2571..2795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2841..3269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3283..3613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3464..3599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3661..3708
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3727..4101
                     /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    4231..4352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4401..4417)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4425..4441
                     /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(4449..4479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4494..4515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4803..5391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5562..6422)
                     /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(6423..6527)
                     /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 agtgcagctg ttgtcagggc ggcaccggtg gcagcaccag caccacactg
     1201 gtcaagtgcg cctgcgagcc ttgcctgtgt aatgtggacc ctagcaaggc catcgaccgg
     1261 aacggcctgt actactgttc tgaggcctgt gccgatggcc acaccggcgg atctaaaggc
     1321 tgtggacaca ccggctgcaa ctgtcacggc tccgcgggtg gctgcgacga taagtgcgga
     1381 tgtgctgtgc cttgtcctgg cggcaccggc tgtagatgta caagcgctag aagcggagcc
     1441 gccgctggcg aacatacaac atgtggatgt ggcgagcact gcggctgcaa tccttgtgcc
     1501 tgtggcagag agggcacacc ttctggcaga gccaacagaa gggccaactg ttcttgtggc
     1561 gccgcttgca attgtgcctc ttgcggctct gctacagccc ctggaggtgg tggaagtggg
     1621 ggaggatcgg gtggatcagc gaacaagagc cagctgtacc ctgacagccc tctgaccgac
     1681 caggacttca accagctgga ccagaccgtg attgaagccg ccagaaggca gctcgtcggc
     1741 agaagattca tcgagctgta cggccctctc ggcagaggca tgcagagcgt gttcaacgac
     1801 atcttcatgg aaagccacga ggccaagatg gacttccagg gcagcttcga caccgaggtg
     1861 gaaagcagca gaagagtgaa ctacacaatc cccatgctgt acaaggactt cgtgctgtac
     1921 tggcgcgacc tggaacagag caaggccctg gacatcccca tcgatttttc cgtggccgcc
     1981 aacgccgcta gagatgtggc ctttctggaa gatcagatga tctttcacgg cagcaaagag
     2041 ttcgacatcc ccggcctgat gaacgtgaag ggcagactga cacacctgat cggcaattgg
     2101 tacgagagcg gcaacgcctt ccaggacatc gtggaagccc ggaacaagct gctggaaatg
     2161 aaccacaacg gcccctacgc tctggtgctg agccctgagc tgtatagcct gctgcacaga
     2221 gtgcacaagg acaccaacgt cctggaaatc gagcacgtgc gcgagctgat taccgccggt
     2281 gtctttcaga gccctgtgct gaagggaaag tccggcgtga tcgtgaacac cggcagaaac
     2341 aacctggacc tggccatcag cgaggacttc gagacagcct atctgggcga agagggcatg
     2401 aatcacccct tccgggtgta cgagacagtg gtgctgcgga ttaagaggcc tgccgccatc
     2461 tgcaccctga tcgatcctga agaaggctcc ggcgactaca aggacgacga cgacaaatga
     2521 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2581 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2641 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2701 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2761 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2821 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3181 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3241 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3301 ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3361 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3421 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3481 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3541 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3601 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     3661 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     3721 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     3781 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     3841 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     3901 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     3961 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     4021 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     4081 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     4141 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4201 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4261 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4321 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4381 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4441 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4501 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4561 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4621 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4681 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4741 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4801 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4861 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4921 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4981 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5041 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5101 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5161 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5221 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5281 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5341 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5401 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5461 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5521 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5581 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5641 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5701 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5761 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5821 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5881 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5941 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6001 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6061 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6121 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6181 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6241 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6301 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6361 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6421 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6481 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6541 aaagtgccac ctgacgtc
//
