LOCUS       sequence-121945-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Terminator quantification vector utilizing Golden Gate Assembly and 
            amilCP. Positive colonies are green and false positives are blue. 
            Terminators are placed between GFP and RFP - induced by arabinose..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  TITLE     Reyna Lab - Terminator Verification Plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5797)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PGR-Blue
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             693..704
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             778..789
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             796..1473
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1518..1589
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1605..1632
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1726..1743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1897..2485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1977..1996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2656..3447,3448..3516))
                     /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     3279..3298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3517..3621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3689..3707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3711..3754)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3843..4721)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4748..5032
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4955..4974
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             5060..5071
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5078..5791
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5108..5136)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5731..5751
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattccgac tgagacctta ttaggcgacc acaggtttgc gtgcaatgga aatttcacac
       61 tgctcaaccg aagtgtaatc cttgttgtga ttggttacat ccagtttgcg gtcaacatag
      121 tgataccctg gcatcttcac aggcttcttt gccttgtaag tagttttaaa ttcacacaaa
      181 tagtgaccgc ctccttctaa cttcagagcc ataaagttgt ttcctagcag cattccatct
      241 cgtgcaaaga gacgctcagt gttgggttcc cagccctgtg tcttcttctg catgacaggt
      301 ccattgggag gaaagttcaa accagagaac ttgacatggt agatgaaaca gttgccttgg
      361 atgctggaat cattgctgac agtacacact gcaccatctt caaagttcat gatcctctcc
      421 catgtatagc cctccgggaa tgactgcttt acatagtcag ggatgtcttc agggtacttg
      481 gtgaatggta tgcttccgta ctgacactgt ggtgataaaa tatcccaagc aaatggcaga
      541 ggtccgccct tggtgacagt gagctttacc gtctgctccc cctcgtaggg cttacctttt
      601 ccatcgcctt cgacctcaaa gtagtgtcca ttgaccgtgc ctgacatata aaccttgtag
      661 gtcatttgtt tagcgatcac actcatctag tatttctcct ctttaattac tagatccaca
      721 cattataggt acaaaaagat gcgaagtcaa tactcttttt ggtctcagcg gctgcagaaa
      781 gaggagaaat actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa
      841 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt
      901 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg
      961 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac
     1021 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt
     1081 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac
     1141 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt
     1201 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt
     1261 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct
     1321 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc
     1381 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa
     1441 cgtgctgaag gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg
     1501 ctactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1561 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1621 ggcctttctg cgtttatata ctagagtcaa ttgaggtaga ggtacacacg cgaactccga
     1681 tagccaattc agagtaataa actgtgataa tcaatgcagt gcttcctcgc tcactgactc
     1741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     3721 aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tgaattgagg tagaggtaca
     3781 cacgcgaact ccgatagcca attcagagta ataaactgtg ataatcaatg catgtactag
     3841 agttatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     3901 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     3961 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     4021 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     4081 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     4141 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     4201 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     4261 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     4321 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     4381 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     4441 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     4501 aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     4561 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     4621 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     4681 gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca
     4741 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
     4801 tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc
     4861 gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc
     4921 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
     4981 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc ataccgtttt
     5041 tttgggctag ctactagaga aagaggagaa atactagatg cgtaaaggag aagaactttt
     5101 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     5161 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     5221 cactactgga aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtgt
     5281 tcaatgcttt gcgagatacc cagatcatat gaaacagcat gactttttca agagtgccat
     5341 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     5401 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     5461 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
     5521 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag
     5581 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     5641 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     5701 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     5761 gattacacat ggcatggatg aactatacaa ataataa
//
