LOCUS       Exported                7614 bp DNA     circular SYN 22-JUN-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 10, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7614
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             907..1293
                     /codon_start=1
                     /label=H2A
                     /note="color: #005493"
                     /translation="MSGRGKQGGKARAKAKSRSSRAGLQFPVGRVHRLLRKGNYAERVG
                     AGAPVYMAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNKLLGKVTIA
                     QGGVLPNIQAVLLPKKTESHKAKSK"
     CDS             1303..2019
                     /codon_start=1
                     /label=Cerulean
                     /note="color: #66ccff"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2029..2382
                     /codon_start=1
                     /label=4EBP1
                     /note="note=""direct strand 
                     translation""translation=""MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPG
                     DYSTTPGGTLFSTTPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPM
                     EASQSHLRNSPEDKRAGGEESQFEMDI"""
                     /note="color: #666666"
                     /translation="MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPGDYSTTPGGTLFST
                     TPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPMEASQSHLRNSPED
                     KRAGGEESQFEMDI"
     CDS             2389..3099
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="color: #e1ff2f"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     LCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLEF
                     LTAAGITEGMNELYK"
     promoter        complement(3166..3184)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    3210..3434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3480..3908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3922..4252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4103..4238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4319..5113
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5287..5408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5457..5473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5481..5497
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5505..5535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5505 .. 5511 / #ffffff / -10
                        2: 5512 .. 5529 / #ffffff
                        3: 5530 .. 5535 / #ffffff / -35"
     protein_bind    5550..5571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5859..6447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6618..7478)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6618 .. 7409 / #ccffcc
                        2: 7410 .. 7478 / #ccffcc / signal sequence
                     Cleavage site after base 7409"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7479..7583)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
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 gcttgcggcc
      901 gccaccatgt ctggtcgtgg caaacaagga ggcaaggccc gcgccaaggc caagtcgcgc
      961 tcgtcccgcg ctggcctcca gttcccggta gggcgagtgc accgcttgct gcgcaaaggc
     1021 aactacgcgg agcgggtggg ggccggcgcg cccgtctaca tggcggcggt cctcgagtac
     1081 ctgaccgccg agatcctgga gctggcgggc aacgcggctc gggacaacaa gaagacgcgc
     1141 atcatccctc gtcacctcca gctggccatc cgcaacgacg aggaactgaa caagctgctg
     1201 ggcaaagtca ccatcgccca gggcggcgtt ttgcctaaca tccaggccgt tctgttacca
     1261 aagaaaaccg aaagccacaa agccaaaagc aaagcggatc ccatggtgag caagggcgag
     1321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1381 aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1501 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1561 tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga cgacggcaac
     1621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc
     1741 atcagtgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcccacttc
     1801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1981 gccgccggga tcactctcgg catggacgag ctgtacaagc gcatgcatat gtccgggggc
     2041 agcagctgca gccagacccc aagccgggcc atccccgcca ctcgccgggt ggtgctcggc
     2101 gacggcgtgc agctcccgcc cggggactac agcacgaccc ccggcggcac gctcttcagc
     2161 accaccccgg gaggtaccag gatcatctat gaccggaaat tcctgatgga gtgtcggaac
     2221 tcacctgtga ccaaaacacc cccaagggat ctgcccacca ttccgggggt caccagccct
     2281 tccagtgatg agccccccat ggaagccagc cagagccacc tgcgcaatag cccagaagat
     2341 aagcgggcgg gcggtgaaga gtcacagttt gagatggaca ttgagctctc taaaggtgaa
     2401 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
     2461 aaattttctg tctccggtga aggtgaaggt gatgctacgt acggtaaatt gaccttaaaa
     2521 ttactctgta ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaggt
     2581 tatggtgttc aatgttttgc tagataccca gatcatatga aacaacatga ctttttcaag
     2641 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     2701 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     2761 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     2821 aactctcaca atgtttacat cactgctgac aaacaaaaga atggtatcaa agctaacttc
     2881 aaaattagac acaacattga agatggtggt gttcaattag ctgaccatta tcaacaaaat
     2941 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc ctatcaatct
     3001 gccttattca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttttgact
     3061 gctgctggta ttaccgaggg tatgaatgaa ttgtacaaat agtaagaatt ctgcagatat
     3121 ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     3181 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3241 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3301 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3361 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3421 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3481 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3541 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3601 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3661 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3721 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3781 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3841 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3901 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3961 ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4081 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4201 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4261 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5281 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5401 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
