LOCUS       Exported                6493 bp ds-DNA     circular SYN 04-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  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..6493
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        279..556
                     /label=chicken beta-actin promoter
     primer_bind     956..1004
                     /label=1108-mIFP-F
     regulatory      977..982
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             983..1942
                     /codon_start=1
                     /product="naturally monomeric infrared fluorescent protein 
                     derived from a truncated bacteriophytochrome (Yu et al., 
                     2015)"
                     /label=mIFP
                     /translation="MSVPLTTSAFGHAFLANCEREQIHLAGSIQPHGILLAVKEPDNVV
                     IQASINAAEFLNTNSVVGRPLRDLGGDLPLQILPHLNGPLHLAPMTLRCTVGSPPRRVD
                     CTIHRPSNGGLIVELEPATKTTNIAPALDGAFHRITSSSSLMGLCDETATIIREITGYD
                     RVMVVRFDEEGNGEILSERRRADLEAFLGNRYPASTIPQIARRLYEHNRVRLLVDVNYT
                     PVPLQPRISPLNGRDLDMSLSCLRSMSPIHQKYMQDMGVGATLVCSLMVSGRLWGLIAC
                     HHYEPRFVPFHIRAAGEALAETCAIRIATLESFAQSQSK"
     primer_bind     complement(1918..1965)
                     /label=1108-mIFP-R
     CDS             1967..2023
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2036..2752
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    2777..3365
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3248..3259)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3408..3529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4057..4645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4816..5676)
                     /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(5677..5781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5807..6262)
                     /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 ccatgtcggt accgctgact acctcagcat tcggccacgc
     1021 gtttctggct aactgtgaac gcgagcagat ccacctggcg ggctccattc agccgcacgg
     1081 tatcctgctg gctgtgaaag agccggacaa cgtggtgatc caggcttcta ttaacgctgc
     1141 ggagttcctg aacaccaact ctgttgttgg ccgtccgctg cgtgacctgg gcggcgatct
     1201 gcctttgcag atcctgccgc acctgaacgg cccgctgcac ctggctccga tgaccctgcg
     1261 ttgtaccgtg ggttctccgc cgcgtcgtgt ggactgtacc attcatcgtc cgtctaacgg
     1321 cggcctgatc gtagaactgg aaccagcaac caagaccact aacattgcgc cggctctgga
     1381 cggtgcgttt catcgtatca cttcttcatc ctccctgatg ggcctgtgtg acgaaaccgc
     1441 gactattatc cgtgagatta ctggctacga ccgtgtgatg gtagtacgtt tcgatgaaga
     1501 gggtaatggc gaaattctgt ccgaacgtcg tcgtgcggac ctggaagcgt tcctgggtaa
     1561 ccgctacccg gcgtctacta ttccgcagat cgctcgtcgc ctgtacgaac ataaccgtgt
     1621 tcgcctgctg gtagatgtga actatactcc ggttccgcta cagccgcgca tcagcccgct
     1681 gaacggtcgt gatctggata tgtccctgtc ttgcctgcgc tctatgtccc cgatccacca
     1741 gaaatacatg caggacatgg gcgttggcgc gaccctggtt tgctctctga tggtgtctgg
     1801 tcgtctgtgg ggtctgatcg cttgccacca ctacgaaccg cgcttcgttc cgttccacat
     1861 tcgcgctgct ggcgaagcgc tggcggaaac ttgtgcgatc cgcatcgcga cgctggagag
     1921 ctttgcacag tctcagtcca aacgaccggt tcacgtggga agtggggcta caaacttctc
     1981 actgctgaaa caggccgggg atgtggagga aaacccagga ccacacgtga ctagtgtgag
     2041 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2101 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2161 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2221 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2281 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2341 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2401 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2461 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2521 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2581 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2641 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2701 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagaattcga
     2761 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2821 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2881 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2941 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3001 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3061 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3121 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3181 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3241 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3301 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3361 cccgcatcga taccgtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     3421 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3481 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3541 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3601 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     3661 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3721 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3781 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca
     3841 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3901 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3961 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4861 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4921 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4981 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5041 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5101 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5161 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5221 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5281 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5341 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5401 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5461 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5521 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5581 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5641 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     5821 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5881 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5941 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6001 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6061 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     6121 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6181 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6241 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6301 cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc
     6361 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     6421 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
     6481 ccgccatgct act
//
