LOCUS       62056_5640        7527 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7527)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1089..1727
                     /codon_start=1
                     /label=mTFP*
                     /note="mTFP* is a basic (constitutively fluorescent) cyan 
                     fluorescent protein published in 2019, derived from 
                     Clavularia sp.."
                     /translation="GVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGTNTINLEVKE
                     GAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFEDKGIVKVKS
                     DISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKGDVKHKLLLE
                     GGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV"
     CDS             2384..3091
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="KGAELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLI
                     CTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANF
                     KIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFV
                     TAAGITLGMDELYK"
     polyA_signal    3127..3351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3397..3825
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3839..4168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4235..5026
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5203..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5373..5389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5397..5413)
                     /label=lac operator
                     /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(5421..5451)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5466..5487)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5775..6360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6534..7391)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7392..7496)
                     /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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtggat ccactcgaga tggcaggctc gaaattagtt gtcgcgacgg
     1021 ataccgcctt cgttccgttc gaatttaaac aggccccaat ggtgagcaag ggcgaggaga
     1081 ccacaatggg cgtaatcaag cccgacatga agatcaagct gaagatggag ggcaacgtga
     1141 atggccacgc cttcgtgatc gagggcgagg gcgagggcaa gccctacgac ggcaccaaca
     1201 ccatcaacct ggaggtgaag gagggagccc ccctgccctt ctcctacgac attctgacca
     1261 ccgcgttcgc ctacggcaac agggccttca ccaagtaccc cgacgacatc cccaactact
     1321 tcaagcagtc cttccccgag ggctactctt gggagcgcac catgaccttc gaggacaagg
     1381 gcatcgtgaa ggtgaagtcc gacatctcca tggaggagga ctccttcatc tacgagatac
     1441 acctcaaggg cgagaacttc ccccccaacg gccccgtgat gcagaagaag accaccggct
     1501 gggacgcctc caccgagagg atgtacgtgc gcgacggcgt gctgaagggc gacgttaagc
     1561 acaagctgct gctggagggc ggcggccacc accgcgttga cttcaagacc atctacaggg
     1621 ccaagaaggc ggtgaagctg cccgactatc actttgtgga ccaccgcatc gagatcctga
     1681 accacgacaa ggactacaac aaggtgaccg tttacgagag cgccgtaccc gcaactccac
     1741 cgacggcatg gacgagctgt acaaggtgag cgccaaatat gtgggctttg acgtcgatct
     1801 gtgggctgcc atcgctaaag agctgaagct ggattacgaa ctgaagccga tggatttcag
     1861 tgggatcatt ccggcactgc aaaccaaaaa cgtcgatctg gcgctggcgg gcattaccat
     1921 caccgacgag cgtaaaaaag cgatcgattt ctctgacggc tactacaaaa gcggcctgtt
     1981 agtgatggtg aaagctaaca ataacgatgt gaaaagcgtg aaagatctcg acgggaaagt
     2041 ggttgctgtg aagagcggta ctggctccgt tgattacgcg aaagcaaaca tcaaaactaa
     2101 agatctgcgt cagttcccga acatcgataa cgcctatatg gaactgggca ccaaccgcgc
     2161 agacgccgtt ctgcacaaca cgccaaacat tctgtacttc atcaaaaccg ccggtaacgg
     2221 tcagttcaaa gcggtaggtg actctctgga agcgcagcaa tacggtattg cgttcccgaa
     2281 aggtagcgac gagctgcgtg acaaagtcaa cggcgcgttg aaaaccctgc gcgagaacgg
     2341 aacttacaac gaaatctaca aaaaatggtt cggtactgaa ccgaaaggcg ctgagctgtt
     2401 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2461 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     2521 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
     2581 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2641 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2701 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2761 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2821 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     2881 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     2941 cggcgacggc cccgtgctgc tgcccgacaa tcactacctg agctaccagt ccgccctgag
     3001 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3061 gatcactctc ggcatggacg agctgtacaa gtaaaagctt aagtttaaac cgctgatcag
     3121 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3181 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3241 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3301 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3361 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3841 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3901 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3961 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4021 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4081 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4141 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4201 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4261 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4321 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4381 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4441 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4501 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4561 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4621 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4681 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4741 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4801 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4861 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4921 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4981 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5041 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5101 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5161 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5221 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5281 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5341 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5401 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5461 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5521 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5581 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5641 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5701 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5761 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5821 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5881 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5941 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6001 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6061 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6121 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6181 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6241 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6301 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6841 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7141 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7501 tttccccgaa aagtgccacc tgacgtc
//
