LOCUS       Exported                7100 bp ds-DNA     circular SYN 05-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7100)
  AUTHORS   U0132803
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 3, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7100
                     /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             926..1705
                     /codon_start=1
                     /label=GalfaS(long)
                     /translation="MGCLGNSKTEDQRNEEKAQREANKKIEKQLQKDKQVYRATHRLLL
                     LGAGESGKSTIVKQMRILHVNGFNGEGGEEDPQAARSNSDGEKATKVQDIKNNLKEAIE
                     TIVAAMSNLVPPVELANPENQFRVDYILSVMNVPDFDFPPEFYEHAKALWEDEGVRACY
                     ERSNEYQLIDCAQYFLDKIDVIKQADYVPSDQDLLRCRVLTSGIFETKFQVDKVNFHMF
                     DVGGQRDERRKWIQCFNDVTAIIFVVASSSYNMVIRED"
     primer_bind     complement(1247..1264)
                     /label=REV_GaSL-113
     primer_bind     1265..1282
                     /label=FWD_GaSL-113
     primer_bind     complement(1307..1333)
                     /label=Rev_GaSL-136
     primer_bind     1334..1356
                     /label=FWD_GaSL-136
     primer_bind     1679..1733
                     /label=FWD_GaSs-244-L-Nluc-L-OH
     primer_bind     1688..1733
                     /label=Fragment.REV
     primer_bind     complement(1688..1705)
                     /label=Vector.FOR
     primer_bind     1705..1733
                     /label=FWD_Gai2-112-L-Nuc-L
     primer_bind     1705..1733
                     /label=FWD_Ga12-133-L-Nluc-L-OH
     primer_bind     1706..1733
                     /label=Fwd_Lnk-NLuc-91Galfai2
     primer_bind     1706..1733
                     /label=FWD_GaQ-124-L-Nuc-L
     primer_bind     1706..1732
                     /label=Fwd-L-NLuc-L
     variation       1706..1723
                     /label=Linker
     primer_bind     1715..1745
                     /label=Fwd_LinkerNLuc
     CDS             1724..2233
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     1724..1748
                     /label=Fwd_NLucfrag_91
     primer_bind     1724..1748
                     /label=Fwd_NLucfrag_122
     primer_bind     complement(2215..2234)
                     /label=Rev_NLucfrag_91
     primer_bind     complement(2215..2233)
                     /label=Rev_NLucfrag_122
     primer_bind     complement(2217..2242)
                     /label=Rev_Nluc-Lnk
     primer_bind     complement(2224..2269)
                     /label=Fragment.FOR
     primer_bind     complement(2224..2263)
                     /label=Rev_GaSs-244-L-Nluc-L-OH
     primer_bind     complement(2224..2252)
                     /label=REV_Gai2-112-L-Nuc-L
     primer_bind     complement(2224..2251)
                     /label=Rev_NLuc-Lnk-Galfai2
     primer_bind     complement(2224..2251)
                     /label=Rev L-Nluc-L
     primer_bind     complement(2224..2251)
                     /label=REV_GaQ-124-L-Nuc-L
     primer_bind     complement(2224..2251)
                     /label=Rev_Ga12-133-L-Nluc-L-OH
     variation       2234..2251
                     /label=Linker
     CDS             2252..2656
                     /codon_start=1
                     /label=GalfaS(long)
                     /translation="NQTNRLQEALNLFKSIWNNRWLRTISVILFLNKQDLLAEKVLAGK
                     SKIEDYFPEFARYTTPEDATPEPGEDPRVTRAKYFIRDEFLRISTASGDGRHYCYPHFT
                     CAVDTENIRRVFNDCRDIIQRMHLRQYELL"
     primer_bind     2252..2269
                     /label=Vector.REV
     polyA_signal    2700..2924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2970..3398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3412..3741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3592..3727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3808..4602
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4776..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4970..4986
                     /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(4994..5024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5039..5060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5348..5933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6104..6964)
                     /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(6965..7069)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ccaccatggg ctgcctcggg aacagtaaga ccgaggacca
      961 gcgcaacgag gagaaggcgc agcgtgaggc caacaaaaag atcgagaagc agctgcagaa
     1021 ggacaagcag gtctaccggg ccacgcaccg cctgctgctg ctgggtgctg gagaatctgg
     1081 taaaagcacc attgtgaagc agatgaggat cctgcatgtt aatgggttta atggagaggg
     1141 cggcgaagag gacccgcagg ctgcaaggag caacagcgat ggtgagaagg caaccaaagt
     1201 gcaggacatc aaaaacaacc tgaaagaggc gattgaaacc attgtggccg ccatgagcaa
     1261 cctggtgccc cccgtggagc tggccaaccc cgagaaccag ttcagagtgg actacatcct
     1321 gagtgtgatg aacgtgcctg actttgactt ccctcccgaa ttctatgagc atgccaaggc
     1381 tctgtgggag gatgaaggag tgcgtgcctg ctacgaacgc tccaacgagt accagctgat
     1441 tgactgtgcc cagtacttcc tggacaagat cgacgtgatc aagcaggctg actatgtgcc
     1501 gagcgatcag gacctgcttc gctgccgtgt cctgacttct ggaatctttg agaccaagtt
     1561 ccaggtggac aaagtcaact tccacatgtt tgacgtgggt ggccagcgcg atgaacgccg
     1621 caagtggatc cagtgcttca acgatgtgac tgccatcatc ttcgtggtgg ccagcagcag
     1681 ctacaacatg gtcatccggg aggacagcgg cggcggcggc agcgtcttca cactcgaaga
     1741 tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg
     1801 aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt
     1861 cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct
     1921 gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga
     1981 tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa
     2041 catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat
     2101 cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc
     2161 cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga
     2221 acgcattctg gcgagcggcg gcggcggcag caaccagacc aaccgcctgc aggaggctct
     2281 gaacctcttc aagagcatct ggaacaacag atggctgcgc accatctctg tgatcctgtt
     2341 cctcaacaag caagatctgc tcgctgagaa agtccttgct gggaaatcga agattgagga
     2401 ctactttcca gaatttgctc gctacactac tcctgaggat gctactcccg agcccggaga
     2461 ggacccacgc gtgacccggg ccaagtactt cattcgagat gagtttctga ggatcagcac
     2521 tgccagtgga gatgggcgtc actactgcta ccctcatttc acctgcgctg tggacactga
     2581 gaacatccgc cgtgtgttca acgactgccg tgacatcatt cagcgcatgc accttcgtca
     2641 gtacgagctg ctctaactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2701 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2761 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2821 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2881 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2941 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3181 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3301 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3421 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3541 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3601 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3661 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3721 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3781 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3841 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3901 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3961 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4021 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4081 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4141 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4201 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4261 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4321 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4381 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4441 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4501 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4561 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4621 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4681 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4741 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4921 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5881 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7081 gaaaagtgcc acctgacgtc
//
