LOCUS       Exported                7770 bp ds-DNA     circular SYN 24-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  AUTHORS   Nico
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7770
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-F
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1790..3325
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     misc_feature    3339..3925
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3506..3523)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3733..3752
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3926..4687
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYMSGGLQKKLEELELD"
     primer_bind     complement(3971..3992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3971..3987)
                     /label=pEGFP_N
     primer_bind     complement(4232..4251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4579..4600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4615..4631)
                     /label=EGFP CR
     primer_bind     4636..4688
                     /label=CNES Fwd
     primer_bind     complement(4640..4692)
                     /label=CNES Rvs
     CDS             4652..4684
                     /codon_start=1
                     /label=NES
                     /translation="LQKKLEELELD"
     primer_bind     complement(4654..4674)
                     /label=NES Colony R
     primer_bind     complement(4762..4781)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4827..4882
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(4881..4900)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5273..5289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5273..5289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5286..5308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5297..5313
                     /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(5321..5351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5366..5387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5446..5641
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5492..5627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5554..5573
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5577..5588)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5647..5781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5684..5703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5738..5757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5849..5866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6020..6608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6100..6119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6779..7639)
                     /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"
     primer_bind     7402..7421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7640..7744)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgct cgagagttcg
     1741 aagccttaag tacgcgtagg cgcgccttaa ttaacgttta aacgctagca tgaagtgcct
     1801 tttgtactta gcctttttat tcattggggt gaattgcaag ttcaccatag tttttccaca
     1861 caaccaaaaa ggaaactgga aaaatgttcc ttctaattac cattattgcc cgtcaagctc
     1921 agatttaaat tggcataatg acttaatagg cacagcctta caagtcaaaa tgcccaagag
     1981 tcacaaggct attcaagcag acggttggat gtgtcatgct tccaaatggg tcactacttg
     2041 tgatttccgc tggtatggac cgaagtatat aacacattcc atccgatcct tcactccatc
     2101 tgtagaacaa tgcaaggaaa gcattgaaca aacgaaacaa ggaacttggc tgaatccagg
     2161 cttccctcct caaagttgtg gatatgcaac tgtgacggat gccgaagcag tgattgtcca
     2221 ggtgactcct caccatgtgc tggttgatga atacacagga gaatgggttg attcacagtt
     2281 catcaacgga aaatgcagca attacatatg ccccactgtc cataactcta caacctggca
     2341 ttctgactat aaggtcaaag ggctatgtga ttctaacctc atttccatgg acatcacctt
     2401 cttctcagag gacggagagc tatcatccct gggaaaggag ggcacagggt tcagaagtaa
     2461 ctactttgct tatgaaactg gaggcaaggc ctgcaaaatg caatactgca agcattgggg
     2521 agtcagactc ccatcaggtg tctggttcga gatggctgat aaggatctct ttgctgcagc
     2581 cagattccct gaatgcccag aagggtcaag tatctctgct ccatctcaga cctcagtgga
     2641 tgtaagtcta attcaggacg ttgagaggat cttggattat tccctctgcc aagaaacctg
     2701 gagcaaaatc agagcgggtc ttccaatctc tccagtggat ctcagctatc ttgctcctaa
     2761 aaacccagga accggtcctg ctttcaccat aatcaatggt accctaaaat actttgagac
     2821 cagatacatc agagtcgata ttgctgctcc aatcctctca agaatggtcg gaatgatcag
     2881 tggaactacc acagaaaggg aactgtggga tgactgggca ccatatgaag acgtggaaat
     2941 tggacccaat ggagttctga ggaccagttc aggatataag tttcctttat acatgattgg
     3001 acatggtatg ttggactccg atcttcatct tagctcaaag gctcaggtgt tcgaacatcc
     3061 tcacattcaa gacgctgctt cgcaacttcc tgatgatgag agtttatttt ttggtgatac
     3121 tgggctatcc aaaaatccaa tcgagcttgt agaaggttgg ttcagtagtt ggaaaagctc
     3181 tattgcctct tttttcttta tcatagggtt aatcattgga ctattcttgg ttctccgagt
     3241 tggtatccat ctttgcatta aattaaagca caccaagaaa agacagattt atacagacat
     3301 agagatgaac cgacttggaa agtaacccgg gggatccgcc cctctccctc ccccccccct
     3361 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3421 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3481 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3541 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3601 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3661 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3721 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3781 gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag
     3841 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3901 acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt
     3961 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4021 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4081 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     4141 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4201 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4261 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4321 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4381 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4441 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4501 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4561 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4621 cggcatggac gagctgtaca tgagcggcgg cctgcagaag aagctggagg agctggagct
     4681 ggactaatgt acaagtaaag cggccggccg ccagcacagt ggtcgatcga ccaatgccct
     4741 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     4801 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     4861 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     4921 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     4981 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     5041 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     5101 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     5161 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     5221 ccgtcgacca gctggtcgac ggtgcaccgt cgaccagctt ggcgtaatca tggtcatagc
     5281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     5461 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5521 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5581 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5641 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6181 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     6241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6421 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7741 cgcgcacatt tccccgaaaa gtgccacctg
//
