LOCUS       sequence-45457-e        6857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  AUTHORS   Van Engelenburg SB, Nahreini T, Palmer AE
  TITLE     FACS-based selection of tandem tetracysteine peptides with improved 
            ReAsH brightness in live cells.
  JOURNAL   Chembiochem. 2010 Mar 1;11(4):489-93.
  PUBMED    20099291
REFERENCE   2  (bases 1 to 6857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Chembiochem."; date: "2010-03-1"; volume: "11(4)"; pages: "489-93"
COMMENT     Sequence Label: pc3xTC20-alphaTubulin
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             938..955
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             983..1000
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             1028..1045
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(2451..2468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2457..2681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2727..3155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2814..2833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3024..3045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3164..3184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3169..3498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3349..3484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3411..3430
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3565..4359
                     /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"
     primer_bind     complement(3619..3638)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4229..4248
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4533..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4570..4589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4624..4643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4703..4719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4703..4719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4716..4738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4727..4743
                     /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(join(4751..4757,4758..4775,4776..4781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4796..4817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4934..4951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5105..5690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5185..5204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5861..6652,6653..6721))
                     /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     6484..6503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6722..6826)
                     /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 aagcttacca tgggcagctt cctgaactgc tgccctggct gctgcatgga
      961 gcctagtcat aagttcctga actgctgccc tggctgctgc atggagcctc gtccgaagtt
     1021 cctgaactgc tgccctggct gctgcatgga gcctggcagc cgtgagtgca tctccatcca
     1081 cgttggccag gctggtgtcc agattggcaa tgcctgctgg gagctctact gcctggaaca
     1141 cggcatccag cccgatggcc agatgccaag tgacaagacc attgggggag gagatgactc
     1201 cttcaacacc ttcttcagtg agacgggcgc tggcaagcac gtgccccggg ctgtgtttgt
     1261 agacttggaa cccacagtca ttgatgaagt tcgcactggc acctaccgcc agctcttcca
     1321 ccctgagcag ctcatcacag gcaaggaaga tgctgccaat aactatgccc gagggcacta
     1381 caccattggc aaggagatca ttgaccttgt gttggaccga attcgcaagc tggctgacca
     1441 gtgcaccggt cttcagggct tcttggtttt ccacagcttt ggtgggggaa ctggttctgg
     1501 gttcacctcc ctgctcatgg aacgtctctc agttgattat ggcaagaagt ccaagctgga
     1561 gttctccatt tacccagcac cccaggtttc cacagctgta gttgagccct acaactccat
     1621 cctcaccacc cacaccaccc tggagcactc tgattgtgcc ttcatggtag acaatgaggc
     1681 catctatgac atctgtcgta gaaacctcga tatcgagcgc ccaacctaca ctaaccttaa
     1741 ccgccttatt agccagattg tgtcctccat cactgcttcc ctgagatttg atggagccct
     1801 gaatgttgac ctgacagaat tccagaccaa cctggtgccc tacccccgca tccacttccc
     1861 tctggccaca tatgcccctg tcatctctgc tgagaaagcc taccatgaac agctttctgt
     1921 agcagagatc accaatgctt gctttgagcc agccaaccag atggtgaaat gtgaccctcg
     1981 ccatggtaaa tacatggctt gctgcctgtt gtaccgtggt gacgtggttc ccaaagatgt
     2041 caatgctgcc attgccacca tcaaaaccaa gcgcagcatc cagtttgtgg attggtgccc
     2101 cactggcttc aaggttggca tcaactacca gcctcccact gtggtgcctg gtggagacct
     2161 ggccaaggta cagagagctg tgtgcatgct gagcaacacc acagccattg ctgaggcctg
     2221 ggctcgcctg gaccacaagt ttgacctgat gtatgccaag cgtgcctttg ttcactggta
     2281 cgtgggtgag gggatggagg aaggcgagtt ttcagaggcc cgtgaagata tggctgccct
     2341 tgagaaggat tatgaggagg ttggtgtgga ttctgttgaa ggagagggtg aggaagaagg
     2401 agaggaatac taactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2641 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2701 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2761 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2821 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2881 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2941 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3001 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3061 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3121 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3181 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3241 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3361 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3421 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3481 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3541 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3601 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3661 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3721 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3781 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3841 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3901 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3961 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4021 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4081 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4141 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4201 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4261 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4321 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4381 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4441 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4501 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4561 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4621 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4681 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4861 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6841 aagtgccacc tgacgtc
//
