LOCUS       Exported                5659 bp ds-DNA     circular SYN 14-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5659
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     regulatory      977..982
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             983..1915
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     primer_bind     complement(1896..1944)
                     /label=0503-iRFP670-R
     misc_feature    1943..2531
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2414..2425)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2574..2695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3982..4842)
                     /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"
     promoter        complement(4843..4947)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4973..5428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
       61 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      121 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      181 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      241 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      301 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      361 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      421 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      481 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      541 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      601 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      661 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      721 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      781 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
      841 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
      901 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
      961 aattggatcc ggtaccgcca ccatggcgcg taaggtcgat ctcacctcct gcgatcgcga
     1021 gccgatccac atccccggca gcattcagcc gtgcggctgc ctgctagcct gcgacgcgca
     1081 ggcggtgcgg atcacgcgca ttacggaaaa tgccggcgcg ttctttggac gcgaaactcc
     1141 gcgggtcggt gagctactcg ccgattactt cggcgagacc gaagcccatg cgctgcgcaa
     1201 cgcactggcg cagtcctccg atccaaagcg accggcgctg atcttcggtt ggcgcgacgg
     1261 cctgaccggc cgcaccttcg acatctcact gcatcgccat gacggtacat cgatcatcga
     1321 gttcgagcct gcggcggccg aacaggccga caatccgctg cggctgacgc ggcagatcat
     1381 cgcgcgcacc aaagaactga agtcgctcga agagatggcc gcacgggtgc cgcgctatct
     1441 gcaggcgatg ctcggctatc accgcgtgat gttgtaccgc ttcgcggacg acggctccgg
     1501 gatggtgatc ggcgaggcga agcgcagcga cctcgagagc tttctcggtc agcactttcc
     1561 ggcgtcgctg gtcccgcagc aggcgcggct actgtacttg aagaacgcga tccgcgtggt
     1621 ctcggattcg cgcggcatca gcagccggat cgtgcccgag cacgacgcct ccggcgccgc
     1681 gctcgatctg tcgttcgcgc acctgcgcag catctcgccc tgccatctcg aatttctgcg
     1741 gaacatgggc gtcagcgcct cgatgtcgct gtcgatcatc attgacggca cgctatgggg
     1801 attgatcatc tgtcatcatt acgagccgcg tgccgtgccg atggcgcagc gcgtcgcggc
     1861 cgaaatgttc gccgacttct tatcgctgca cttcaccgcc gcccaccacc aacgctaaga
     1921 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     1981 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2041 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2101 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2161 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2221 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2341 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2401 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2461 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2521 cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata
     2581 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2641 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2701 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     2761 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     2821 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     2881 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2941 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc attaatgaat
     3001 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     4981 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5041 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5101 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5161 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5221 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5281 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5341 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5401 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     5461 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact
     5521 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     5581 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
     5641 attaacccgc catgctact
//
